Search Images Maps Play YouTube News Gmail Drive More »
Sign in
Screen reader users: click this link for accessible mode. Accessible mode has the same essential features but works better with your reader.

Patents

  1. Advanced Patent Search
Publication numberCN101793826 B
Publication typeGrant
Application numberCN 200910246173
Publication date13 Apr 2016
Filing date7 Jun 2005
Priority date7 Jun 2004
Also published asCN100583434C, CN101002322A, CN101793826A, CN102680440A, CN103884698A, CN103884698B, EP1754257A2, EP1754257A4, EP1754257B1, EP2701194A2, EP2701194A3, EP2752874A1, EP2933630A2, EP2933630A3, US7307802, US7588672, US7906072, US8048378, US8512640, US8721968, US8926905, US9234237, US9663821, US20060006067, US20080088952, US20090294703, US20100320364, US20120035080, US20120264226, US20140011204, US20140024559, US20150203896, US20160194685, WO2005121864A2, WO2005121864A3, WO2005121864A9
Publication number200910246173.0, CN 101793826 B, CN 101793826B, CN 200910246173, CN-B-101793826, CN101793826 B, CN101793826B, CN200910246173, CN200910246173.0
InventorsMA厄金, GR费瑟, B克拉克森, CG西泽, N斯威兹
Applicant先锋生物科技股份有限公司
Export CitationBiBTeX, EndNote, RefMan
External Links: SIPO, Espacenet
用于微流体器件的光学透镜系统和方法 An optical lens system and method for microfluidic devices translated from Chinese
CN 101793826 B
Abstract  translated from Chinese
一种给来自微流体器件(205)的一种或多种选定的荧光指示成像的装置。 And one from one kind of microfluidic device (205) of one or more selected fluorescence imaging apparatus to indicate. 该装置包括成像通路,该成像通路耦合到至少一个微流体器件(205)中的至少一个反应室。 The apparatus includes an imaging path, the imaging path coupled to at least one microfluidic device (205) in at least one reaction chamber. 该成像通路可以传输从至少一个微流体器件(205)的至少一个反应室中的一个或多个样品那儿获得的一个或多个荧光发射信号。 One or more fluorescent emission signals can be transferred from the imaging path at least one microfluidic device (205) at least one reaction chamber where one or more sample obtained. 该反应室具有一定的反应室大小,该反应室大小的特征在于其实际空间维度与成像通路正交。 The reaction chamber has a certain size of the reaction chamber, the reaction chamber characterized in that the size of the actual spatial dimension orthogonal to the imaging path. 该装置还包括耦合到成像通路的光学透镜系统(210,212)。 The apparatus further includes an imaging path coupled to an optical lens system (210, 212). 该光学透镜系统适用于传输与该反应室相关联的一个或多个荧光信号。 The optical lens system suitable for a transmission associated with the reaction chamber or a plurality of fluorescent signals.
Claims(31)  translated from Chinese
1. 一种给微流体器件成像的方法,所述方法包括: 用热控制器使微流体器件的不止96个室热循环; 通过光学通道传输所述微流体器件的空间区域的像,所述微流体器件的空间区域与热循环的不止96个室有关; 使第一透镜系统耦合到所述光学通道的第一部分,所述第一透镜系统具有第一光学特性; 使第二透镜系统耦合到所述光学通道的第二部分,所述第二透镜系统具有第二光学特性;以及通过图像检测设备利用与图像检测设备有关的图像检测空间区域来拍摄所述微流体器件的空间区域的图像,其中所述微流体器件的空间区域的面积除以所述图像检测空间区域的面积在1至1/0. 5的范围内。 CLAIMS 1. A microfluidic device for imaging, the method comprising: hot chamber controller causes more than 96 thermal cycles of the microfluidic device; the optical channel transport through the microfluidic device as the space region, the chamber 96 than the space region and a thermal cycling microfluidic device dependent; the first lens system is coupled to a first portion of said optical path, said first lens system having a first optical characteristic; lens system is coupled to the second said second portion of the optical path, said second lens system having a second optical characteristic; and the relevant spatial image detection area to capture an image of the spatial region of the microfluidic device by use of the image detection apparatus and an image detecting device, wherein the area of the space region microfluidic device divided by the image detection spatial region is an area in the range of 1 to 1/0. 5.
2. 如权利要求1所述的方法,其特征在于,所述微流体器件的空间区域的面积除以所述图像检测空间区域的面积大约等于一(1)。 2. The method according to claim 1, characterized in that the area of the space region of the microfluidic device divided by the area of the image detection spatial region is approximately equal to one (1).
3. 如权利要求1所述的方法,其特征在于,所述微流体器件的空间区域的面积除以所述图像检测空间区域的面积在1/0. 99至1/0. 5的范围内。 3. The method according to claim 1, characterized in that the area of the space area of the microfluidic device region divided by the image detection spatial region is in the range of 1/0. 1/0 to 99. 5 .
4. 如权利要求3所述的方法,其特征在于,所述微流体器件的空间区域的面积除以所述图像检测空间区域的面积在1/0. 88至1/0. 5的范围内。 4. The method according to claim 3, characterized in that the area of the space area of the region of the microfluidic device divided by the image detection spatial region is 1/0. Within the range of from 88 to 1/0. 5 .
5. 如权利要求1所述的方法,其特征在于,所述图像检测空间区域大约是30. 7mmX30. 7mm 或更大。 5. The method according to claim 1, wherein the image detection spatial region is about 30. 7mmX30. 7mm or more.
6. 如权利要求1所述的方法,其特征在于,所述图像检测空间区域大约是27. 6mmX 27. 6mm 或更大。 6. The method according to claim 1, wherein the image detection spatial region is about 27. 6mmX 27. 6mm or more.
7. 如权利要求1所述的方法,其特征在于,所述图像检测空间区域包括大约2048X2048个像素区域。 7. The method according to claim 1, wherein the image detection spatial region comprises a region of approximately 2048X2048 pixels.
8. 如权利要求1所述的方法,其特征在于,所述图像检测空间区域包括大约2048 X 2048个像素区域,每一个像素区域具有约为15 μ mX 15 μ m或更大的空间维度。 8. The method according to claim 1, wherein the image detection spatial region comprises about 2048 X 2048 pixel areas, each pixel area having approximately 15 μ mX 15 μ m or more spatial dimensions.
9. 如权利要求8所述的方法,其特征在于,至少两个像素区域与至少一个室相关。 9. The method according to claim 8, characterized in that the at least two pixel regions associated with the at least one chamber.
10. 如权利要求9所述的方法,其特征在于,至少九个像素区域与至少一个室相关。 10. The method according to claim 9, characterized in that at least nine pixel region associated with the at least one chamber.
11. 如权利要求10所述的方法,其特征在于,不止20个像素区域与至少一个室相关。 11. The method according to claim 10, characterized in that, more than 20 pixel region associated with the at least one chamber.
12. -种给微流体器件成像的方法,所述方法包括: 使微流体器件的至少确定数目的室维持在确定的温度范围中; 通过光学通道传输所述微流体器件的空间区域的像; 使第一透镜系统耦合到所述光学通道的第一部分,所述第一透镜系统具有第一光学特性; 使第二透镜系统耦合到所述光学通道的第二部分,所述第二透镜系统具有第二光学特性;以及在使所述确定数目的室维持在确定的温度范围中的同时,在小于1分钟的时间帧内通过图像检测设备利用与图像检测设备有关的图像检测空间区域给和所述微流体器件的所述确定数目的室相关的空间区域拍摄图像,如形成于所述图像检测设备上那样在哪儿拍摄所述微流体器件的空间区域的图像基本上不受缝合和/或扫描过程的影响,其中,所述图像检测空间区域具有和所述微流体器件的空间区域大约相等或更小的尺寸。 12. - kind to the method of imaging microfluidic device, the method comprising: determining at least microfluidic device number of chambers maintained at the determined temperature range; passage through an optical transmission image of the spatial region of the microfluidic device; the first lens system coupled to the first portion of the optical path, the first lens system having a first optical characteristic; the second lens system coupled to a second portion of said optical path, said second lens system having a second optical characteristic; and determining the number of said chamber is maintained at determined temperature range at the same time, in a time frame of less than one minute by the use of an image detection device and image detection apparatus related to the image detection spatial region and the the microfluidic device of said chamber to determine the spatial region associated with the number of the captured image, as formed on the image sensing apparatus as where the captured image of the spatial region of the microfluidic device substantially unaffected by stitching and / or scanning could influence the process wherein the image detection spatial region having the spatial region of the microfluidic device of about the same size or smaller.
13. 如权利要求12所述的方法,还包括: 在大于10毫秒的时间帧内,通过图像检测设备利用图像检测空间区域,给和所述微流体器件中至少确定数目的室相关的空间区域拍摄图像。 13. The method of claim 12, further comprising: greater than 10 ms time frame, by the image detecting device using an image detection spatial region, and the microfluidic device to determine at least a number of chambers in the relevant spatial region Take the picture.
14. 如权利要求12所述的方法,其特征在于,拍摄所述微流体器件的空间区域的图像是在小于30秒的时间帧内发生的。 14. The method according to claim 12, characterized in that the spatial region of the captured image of the microfluidic device is less than 30 seconds at the time of occurrence of the frame.
15. 如权利要求12所述的方法,其特征在于,拍摄所述微流体器件的空间区域的图像是在小于15秒的时间帧内发生的。 15. The method according to claim 12, characterized in that the spatial region of the captured image of the microfluidic device is less than 15 seconds at the time of occurrence of the frame.
16. 如权利要求12所述的方法,其特征在于,在所述小于1分钟的时间帧内,对所述确定数目的室进行同步拍摄。 16. The method according to claim 12, characterized in that, in the time frame of less than one minute, for determining the number of chambers sync shooting.
17. 如权利要求12所述的方法,其特征在于,所述确定的温度范围是2C。 17. The method according to claim 12, characterized in that said determined temperature range is 2 C.
18. 如权利要求12所述的方法,其特征在于,所述确定的温度是与PCR过程相关的循环温度环境的一部分。 18. The method of claim 12, wherein said determined temperature is part of the PCR process, the cycle the temperature of the environment.
19. 如权利要求12所述的方法,其特征在于,所述确定数目的室是至少96个或更多。 19. The method of claim 12, wherein said determining number of chambers is at least 96 or more.
20. 如权利要求12所述的方法,还包括: 在小于1分钟的时间帧内,通过图像检测设备利用图像检测空间区域,给和所述微流体器件中至少确定数目的室相关的空间区域拍摄至少第二张图像,在哪儿拍摄所述微流体器件的空间区域的第二张图像代表了荧光发射。 20. The method of claim 12, further comprising: a time frame of less than one minute, by the image detecting device using an image detection spatial region, and the microfluidic device to determine at least a number of chambers in the relevant spatial region at least the second shooting images in the space where the shooting area of the microfluidic device of the second image represents the fluorescence emission.
21. -种给包括多个处理区域的微流体器件成像的装置,所述多个处理区域包含从Μ 种样品中选出的至少一种样品以及从Ν种试剂中选出的至少一种试剂,所述装置包括: 耦合到微流体器件的热控制器,所述热控制器适用于使所述微流体器件的多个处理区域热循环; 照明系统,耦合到所述微流体器件并适用于用电磁辐射来照射所述微流体器件; 成像系统,沿着一成像通道耦合到所述微流体器件并适用于接收从热循环的所述多个处理区域那儿发出的电磁辐射;以及耦合到所述成像系统的检测器, 其中,所述检测器利用与检测器有关的图像检测空间区域给所述微流体器件的空间区域拍摄图像, 其中,所述图像检测空间区域具有和所述微流体器件的空间区域大约相等或更小的尺寸, 其中,所述成像系统包括耦合到所述成像通道的第一部分的第一透镜系统,所述第一透镜系统具有第一光学特性,其中,所述成像系统包括耦合到所述成像通道的第二部分的第二透镜系统,所述第二透镜系统具有第二光学特性。 21. - kind to the imaging device comprising a plurality of microfluidic processing area of the apparatus, the plurality of sample processing region comprises at least one species selected from Μ sample and at least one kind of agent selected from the Ν reagent , the apparatus comprising: a controller coupled to the heat of the microfluidic device, the controller is adapted to heat the microfluidic device so that a plurality of processing regions thermal cycle; lighting system coupled to the microfluidic device and adapted to with electromagnetic radiation to irradiate the microfluidic device; imaging systems, imaging along a channel coupled to the microfluidic device and adapted to receive electromagnetic radiation from the plurality of processing regions where thermal cycling issued; coupled to the said imaging system detector, wherein the detector is a detector associated with the use of an image detection spatial region to a spatial region of the micro fluidic devices of a captured image, wherein the image detection spatial region of the microfluidic device and having the spatial region is approximately equal or smaller size, wherein the imaging system comprises a first lens system coupled to the first portion of the imaging channel, said first lens system having a first optical characteristic, wherein said imaging the imaging system includes a channel coupled to the second portion of the second lens system, said second lens system having a second optical characteristic.
22. 如权利要求21所述的装置,其特征在于,所述微流体器件是一种弹性微流体器件。 22. The apparatus according to claim 21, wherein said microfluidic device is an elastomeric microfluidic device.
23. 如权利要求21所述的装置,其特征在于,所述处理区域是封闭的反应室。 23. The apparatus according to claim 21, characterized in that the processing region is a closed reaction chamber.
24. 如权利要求23所述的装置,其特征在于,所述封闭的反应室包括每平方厘米有大约250个流体隔离的反应室或更大的反应室密度。 24. The apparatus according to claim 23, characterized in that said chamber comprises a closed reactor per square centimeter approximately 250 fluidly isolated from the reaction chamber or reaction chamber of greater density.
25. 如权利要求21所述的装置,其特征在于,从所述多个处理区域中发出的电磁辐射是荧光信号。 25. The apparatus according to claim 21, characterized in that the electromagnetic radiation emitted from the plurality of processing areas is a fluorescent signal.
26. 如权利要求21所述的装置,其特征在于,所述多个处理区域以大约100纳升或更小的体积为特征。 26. The apparatus according to claim 21, wherein said plurality of processing zone of about 100 nanoliters or smaller volume characteristics.
27. 如权利要求26所述的装置,其特征在于,所述多个处理区域以大约10纳升或更小的体积为特征。 27. The apparatus according to claim 26, wherein said plurality of processing zone of about 10 nanoliter or smaller volumes characterized.
28. 如权利要求26所述的装置,其特征在于,所述多个处理区域以每平方厘米有250个区域或更大的阵列密度为特征。 28. The apparatus according to claim 26, wherein said plurality of processing regions per square centimeter or larger region 250 is characterized by a density arrays.
29. 如权利要求26所述的装置,其特征在于,所述多个处理区域以每平方厘米有1000 个区域或更大的阵列密度为特征。 29. The apparatus according to claim 26, wherein said plurality of processing regions per square centimeter or larger region 1000 is characterized by a density arrays.
30. 如权利要求21所述的装置,其特征在于,所述多个处理区域适用于支持蛋白质结晶过程。 30. The apparatus according to claim 21, characterized in that said plurality of processing regions adapted to support a protein crystallization process.
31. 如权利要求21所述的装置,其特征在于,所述电磁辐射是光辐射。 31. The apparatus according to claim 21, characterized in that said electromagnetic radiation is light radiation.
Description  translated from Chinese
用于微流体器件的光学透镜系统和方法 An optical lens system and method for microfluidic devices

[0001] 本发明专利申请是申请日为2005年6月7日、申请号为200580025595. 5、名称为"用于微流体器件的光学透镜系统和方法"的发明专利申请的分案申请。 [0001] The present invention patent application was filed on June 7, 2005, Application No. 200580025595.5, entitled "optical lens systems and methods for microfluidic devices," the divisional application invention patent applications.

[0002] 有关申请的香叉参照 [0002] Referring to the application of incense fork

[0003] 本申请要求2004年6月7日提交的美国临时申请60/578, 106的优先权。 [0003] This application claims the United States June 7, 2004 to submit the priority provisional application 60/578, 106.

技术领域 TECHNICAL FIELD

[0004] 本发明一般设及微流体技术。 [0004] The present invention generally located and microfluidics. 特别是,本发明提供用于给微流体器件的室内的(例如悬浮在流体体积中的)一个或多个实体成像的方法和系统。 In particular, the present invention is to provide a chamber for microfluidic devices (for example, suspended in a fluid volume of) one or more entities imaging methods and systems. 更具体地讲,运种成像方法和系统利用了与微流体器件中的一个或多个实体有关的巧光信号指示。 More specifically, the image forming method and system of transport use with microfluidic devices in one or more entities associated clever optical signal indicator. 仅作为示例,通过使用禪合到微流体器件的巧光、化学发光和生物发光读出器,来应用微流体方法和系统技术,但是将会认识到本发明具有更广的应用范围。 Only as an example, by using a microfluidic device bonded to the Zen clever light, chemiluminescence and bioluminescence reader to apply microfluidic methods and systems technology, it will be recognized that the invention has a wider range of applications. 阳〇化]经共同努力已开发并制造出各种微流体系统,来执行化学和生物化学的各项分析与综合。 Yang billion of the] joint efforts have been developed and manufactured a variety of microfluidic systems to perform various chemical and biochemical analysis and synthesis. 运些系统已被开发成用于各项制备和分析应用。 These systems have been shipped to developed for the preparation and analysis applications. 制造运种微尺寸器件的目标源自常规大尺度分析与综合(运通常很笨重且不太有效)的小型化过程所实现的诸多重要益处。 Target species transported manufacturing microscale devices from conventional large-scale analysis and synthesis of a number of important benefits (shipped usually very cumbersome and less effective) miniaturization of process implementation. 通过使用运些微流体系统,便实现了时间大幅减少、成本更低、空间分配更有效等诸多益处。 By using slightly fluid transportation system, it will significantly reduce the time to achieve a lower cost, more efficient space allocation, and many other benefits. 对于使用运些微流体器件的自动化系统而言,附带的好处可W包括人工操作更少。 For use slightly fluid transport device automation system, fringe benefits may include manual W less. 自动化系统还减少了操作人员的过失W及其它和操作人员有关的局限。 Automated system also reduces the operator's negligence W and other limitations and operating personnel. 已经提出,微流体器件可用于各种应用,其中包括毛细电泳、气体层析和细胞分离。 It has been proposed, the microfluidic device can be used for various applications, including capillary electrophoresis, gas chromatography and cell separation.

[0006] 适于进行核酸扩增过程的微流体器件有可能有广泛的应用。 [0006] adapted to perform a nucleic acid amplification processes have microfluidic device may have wide application. 例如,运种器件作为一种分析工具,可W用于确定在样品中是否存在特定的祀核酸。 For example, such devices are shipped as an analytical tool that can be used to determine whether the presence of W worship particular nucleic acid in a sample. 利用微流体器件作为分析工具的示例包括: Example use microfluidic device as an analytical tool include:

[0007] •鉴定过程(例如,亲子鉴定和法医学应用); [0007] • Identification of process (for example, paternity and forensic applications);

[0008] •检测和表征与特定疾病或遗传病相关联的特定核酸; [0008] • detection and characterization of nucleic acid associated with a particular disease or a particular genetic disease associated;

[0009] •检测与特定药物行为相关联的基因表达分布型/序列(例如药物遗传学,即选择对特定遗传概况相容的/特别有效的/无害的药物);W及 [0009] • particular drug detection and gene expression profiles associated with the behavior pattern / sequence (such as pharmacogenetics, namely selection of a particular genetic profile compatible / particularly effective / harmless drugs); W and

[0010] •进行基因型分析和基因表达分析(例如,差别基因表达研究)。 [0010] • genotyping and gene expression analysis (for example, differential gene expression studies) conducted.

[0011] 或者,该器件可W按制备方式用于核酸扩增,从而产生足W进行进一步分析的水平的扩增产物。 [0011] Alternatively, the device can be prepared by W mode for nucleic acid amplification, to generate enough W for further analysis of the level of amplification product. 运些分析方法的示例包括扩增产物的测序、细胞分型、DNA指纹鉴定等。 Some examples of transport analysis methods include amplification product sequenced cell typing, DNA fingerprinting and the like. 扩增产物也可用于各种遗传工程应用。 Amplified product can also be used for a variety of genetic engineering applications. 运些遗传工程应用包括(但不限于)生产所需蛋白质产物,运是通过将扩增产物插入载体,然后用该载体转化细胞使其产生所需蛋白质产物完成的。 Transport some genetic engineering applications include (but are not limited to) the production of the desired protein product, is transported by the amplified product inserted into the vector, and then the cells transformed with the vector to produce the desired protein product is completed.

[0012] 尽管有运些潜在的应用,适用于从运种微流体器件中收集并处理成像数据(例如,巧光数据)的成像系统(也被称为读出器)具有各种缺点。 [0012] Although there are some potential applications shipped for species collected from transport microfluidic device and process imaging data (for example, clever optical data) imaging system (also known as a reader) have various disadvantages. 一些常规的读出器按扫描模式工作,其中激光束在微流体器件上进行光栅扫描。 Some conventional reader work by scanning mode, wherein the laser beam on the microfluidic devices raster scan. 在其它运样的系统中,器件被平移,或者激光和器件同时平移。 In other kind of transport system, the device is translated, or laser devices and simultaneously translated. 运些扫描器按照和激光源/器件的光栅扫描相关联的有序方式, 从微流体器件中存在的反应室中收集巧光数据。 These scanners shipped in an orderly manner and associated raster scanning laser source / device exists from the microfluidic device in the reaction chamber to collect clever optical data. 其它常规的扫描器按缝合模式工作,按顺序地给小区域(例如,尺寸小于Imm2的区域)成像,并且将运些小图像缝合到一起,从而形成测试中的微流体器件的图像。 Other conventional scanner operation by stitch pattern, sequentially to a small area (e.g., area size smaller than Imm2) imaging, and will be transported to some small images stitched together to form a test image of the microfluidic device.

[0013] 扫描和缝合模式都有缺点。 [0013] scan and stitch pattern has drawbacks. 例如,两种类型的系统都工作在相对低的系统频率下, 而系统频率作为时间的函数正比于成像面积。 For example, both types of systems operate at a relatively low frequency system, and the system frequency as a function of time is proportional to the imaging area. 常规系统的工作频率在每分钟l-20cm 2的量级。 Conventional system frequency per minute in the order of 2 l-20cm. 对于某些感兴趣的试验,比如蛋白质热量测定和核酸扩增,通常需要大于约每分钟l-20cm 2的系统频率,W便对微流体器件的反应器皿中发生的巧光过程进行成像。 For some tests of interest, such as proteins and nucleic acid amplification calorimetry, usually greater than the system frequency of about l-20cm 2 per minute, W will react to the clever optical process vessels in the microfluidic device for imaging. 常规的扫描和缝合系统无法满足运些性能目标。 Conventional scanning and suturing system can not meet the transportation of these performance goals. 除了使系统通量放缓W外,运些扫描和缝合系统还会限制利用某些试验(例如,实时PCR的性能)的可能性。 In addition to slowing down the system throughput W, the scanning and transport some suturing system will restrict the use of certain tests (for example, the performance of real-time PCR) possibilities.

[0014] 因此,本领域需要改进的方法和系统,来对微流体器件的反应室中的流体体积中悬浮的一个或更多的实体进行成像。 [0014] Thus, the need in the art for improved methods and systems for imaging entity to one or more of the reaction chamber microfluidic device in fluid volume suspended.

发明内容 SUMMARY

[0015] 根据本发明,提供了用于微流体系统的技术。 [0015] The present invention provides a technique for microfluidic systems. 特别是,本发明提供了用于给微流体器件的反应室中的流体体积中悬浮的一个或多个实体成像的方法和系统。 In particular, the present invention provides for the reaction chamber to the microfluidic device in fluid volume in the suspension method and system for imaging one or more entities. 更具体地讲,本发明的成像方法和系统利用了与微流体器件中的一个或多个实体相关联的巧光信号指示。 More specifically, the imaging method and system of the present invention utilize microfluidic device with one or clever optical signal indicating a plurality of associated entities. 仅仅作为示例,通过使用和微流体器件相禪合的巧光、化学发光和生物发光读出器,来应用微流体方法和系统技术,但是将会认识到本发明具有更广的应用范围。 Just as an example, by using a microfluidic device and meditation combined with clever light, chemiluminescence and bioluminescence reader to apply microfluidic methods and systems technology, it will be recognized that the invention has a wider range of applications.

[0016] 在特定的实施例中,本发明提供了一种给微流体器件中的一个或多个选出的巧光指示成像的装置。 [0016] In a particular embodiment, the present invention provides a clever optical imaging device for indicating to the microfluidic device in one or more selected. 该装置包括成像通道,禪合到至少一个微流体器件中的至少一个反应室。 The apparatus includes an imaging channel, Zen bonded to at least one microfluidic device in at least one reaction chamber. 该成像通道用于传输从上述至少一个微流体器件的至少一个反应室中的一个或多个样品那儿获得的一个或多个巧光发射信号。 Forming a channel for the transmission of at least one from said at least one reaction chamber of the microfluidic device of one or more sample to get there, one or more light emitting coincidence signal. 该反应室具有一定的大小,其实际空间维度与成像通道正交。 The reaction chamber has a certain size, and the actual spatial dimension orthogonal to the imaging channel. 该装置还包括禪合到成像通道的光学透镜系统。 The apparatus also includes a Zen bonded to the optical lens system imaging channel. 该光学透镜系统用于传输与该反应室有关的一个或多个巧光信号。 The optical lens system for one or more clever optical signal transmission and the associated reaction chamber.

[0017] 在另一个特定的实施例中,提供了一种用于给微流体器件的至少一个反应室中的一个或多个选出的巧光指示成像的方法。 [0017] In another particular embodiment, at least one reaction chamber provides a microfluidic device is used for the one or clever method of imaging a plurality of light indication selected. 该方法包括:沿禪合到至少一个反应室的成像通道,传输从至少一个微流体器件的至少一个反应室内的一个或多个样品那儿获得的一个或多个巧光发射信号。 The method comprising: Zen along bonded to at least one reaction chamber imaging channel, one or more clever optical transmission obtained from the reaction of at least one of the at least one microfluidic device of one or more sample chamber where the transmitted signal. 上述至少一个反应室具有一定的大小,其实际空间维度与成像通道正交。 It said at least one reaction chamber having a certain size, and the actual spatial dimension orthogonal to the imaging channel. 该方法还包括:通过禪合到成像通道的光学透镜系统,传输与该反应室相关联的一个或多个巧光发射信号。 The method further comprising: Zen bonded to the optical lens system of the imaging channel, a transmission associated with the reaction chamber or more clever optical transmission signal. 该光学透镜系统用于将实际空间尺寸的大小减小到确定的水平。 The optical lens system is used to reduce the size of the actual spatial dimension to a determined level.

[0018] 在本发明的另一个特定实施例中,提供了一种用于给微流体器件的一个或多个反应室内的一个或多个指示成像的系统。 [0018] In another particular embodiment of the present invention, there is provided a microfluidic device used for the reaction chamber of the one or more instructions of one or more imaging systems. 该系统包括光学通道,该光学通道能够传输微流体器件的空间区域的一部分的一个或多个图像。 The system includes an optical path, the optical channels can transmit a part of a spatial region of the microfluidic device or a plurality of images. 在一个实施例中,上述微流体器件的空间区域的那部分具有第一尺寸。 In one embodiment, the portion of the spatial region of the microfluidic device having the above-described first dimension. 该系统还包括第一透镜系统,它禪合到上述光学通道的第一部分。 The system further includes a first lens system, it Zen bonded to the first portion of the optical path. 该第一透镜系统具有第一光学特性。 The first lens system having a first optical characteristic. 该系统还包括第二透镜系统,它禪合到上述光学通道的第二部分。 The system further includes a second lens system, which Zen bonded to the second portion of said optical channels. 第二透镜系统具有第二光学特性。 A second lens system having a second optical characteristic. 该系统另外包括检测器,该检测器禪合到上述光学通道的第=部分。 The system additionally includes a detector, the detector close to the first Zen = portion of the optical channel. 该检测器可用于给上述空间区域的那部分拍摄一个或多个图像。 That portion of the detector can be used to capture said spatial region of one or more images. 此外,该检测器适用于拍摄一个或多个图像。 In addition, the detector is adapted to capture one or more images. 在该检测器处,上述一个或多个图像具有确定的尺寸,约为第一尺寸或更小。 At the detector, the one or more images having a determined size, about a first size or smaller.

[0019] 在可选实施例中,提供了一种给微流体器件的一个或多个反应室的一个或多个指示成像的方法。 [0019] In an alternative embodiment, there is provided a microfluidic device to one or a plurality of reaction chambers of the methods of imaging one or more directions. 该方法包括:沿光学通道,传输微流体器件的空间区域的一部分的一个或多个图像。 The method comprises: along the optical path, a part of a spatial region of the microfluidic device transmitting one or more images. 上述微流体器件的空间区域的那部分具有第一尺寸。 That part of the space region of the microfluidic device having a first dimension. 该方法还包括:将第一透镜系统禪合到光学通道的第一部分。 The method further comprising: a first lens system Zen bonded to a first portion of the optical path. 第一透镜系统具有第一光学特性。 A first lens system having a first optical characteristic. 该方法另外包括:将第二透镜系统禪合到光学通道的第二部分。 The method further comprises: a second lens system Zen bonded to the second portion of the optical path. 第二透镜系统具有第二光学特性。 A second lens system having a second optical characteristic. 此外,该方法包括:用检测器给上述空间区域的那部分拍摄一个或多个图像。 Moreover, the method comprising: one or more images captured by the detector in the space to the area of that part. 该检测器禪合到光学通道的第=部分,并且在检测器处,一个或多个图像具有确定的尺寸,约为第一尺寸或更小。 The detector is bonded to the first Zen = portion of the optical path, and at the detector, having one or more images to determine the size of about a first size or smaller.

[0020] 在另一个可选实施例中,提供了一种给微流体器件成像的方法。 [0020] In a further alternative embodiment, there is provided a microfluidic device to the image forming method. 该方法包括:在小于一分钟的时间范围内,通过使用图像检测空间区域,给和至少确定数目的微流体器件反应室相关联的空间区域拍摄图像。 The method comprises: in the range of less than a minute, by using an image detection spatial region, and to determine at least the spatial region of the microfluidic device number associated with the reaction chamber of the captured image. 在特定的实施例中,上述空间区域的图像拍摄基本上不受缝合和/或扫描过程的影响。 In a particular embodiment, the above-described image-capturing region of space substantially unaffected by impact of the suture and / or scanning process.

[0021] 在另一个可选实施例中,提供了一种给微流体器件的一个或多个选出的巧光指示成像的装置。 [0021] In an alternative embodiment, there is provided a clever light indicates imaging apparatus to one or more microfluidic devices selected. 该装置包括成像通道,该成像通道禪合到至少一个微流体器件的至少一个反应室。 The apparatus includes an imaging channel, the imaging channel Zen bonded to at least one of at least one reaction chamber of the microfluidic device. 该成像通道用于传输从上述至少一个微流体器件的至少一个反应室内的一个或多个样品那儿获得的一个或多个巧光发射信号。 The imaging channel for transmitting a transmission signal from at least one of at least one reaction chamber or a plurality of samples obtained where the above-described microfluidic device or a plurality of optical coincidence. 该装置还包括滤光器件,它禪合到成像通道(用于传输一个或多个发射信号)的第一空间部分。 The apparatus further comprises a filter device, it is bonded to the Zen imaging channels (for transmitting one or more transmit signals) of the first space part. 滤光器件用于传输一个或多个巧光发射信号的选定光谱带宽,还用于将与一个或多个巧光发射信号相关联的一种或多种色差处理到确定的程度。 Optical device for transmitting one or more light emitting skillfully selected spectral bandwidth of the signal, but also for one or more of the color difference signals associated with one or more light emitting clever handling to the extent determined.

[0022] 在特定的实施例中,提供了一种在弹性微流体器件中分析各种处理的方法。 [0022] In a particular embodiment, there is provided a method of analyzing various processes in elastomeric microfluidic devices. 该方法包括:在不到一分钟的时间周期内,拍摄至少96个反应室的图像。 The method comprising: in less than a minute period of time, the captured image at least 96 of the reaction chamber. 在一个实施例中,运至少96个反应室处于彼此之间的流体隔离状态。 In one embodiment, the transport at least 96 of the reaction chamber in fluid isolation between each other. 该方法还包括处理该图像。 The method further includes processing the image.

[0023] 在另一个特定实施例中,提供了一种用于给包括多个处理位置的微流体器件成像的装置。 [0023] In another particular embodiment, there is provided an apparatus for processing to a plurality of positions comprising microfluidic device imaging. 上述多个处理位置包含选自M个样品的至少一个样品W及选自N种试剂的至少一种试剂。 Processing said plurality of selected positions comprises at least M samples and a sample W is selected from N-reagent at least one reagent. 该装置包括照明系统,该照明系统禪合到微流体器件并用电磁福射照亮该微流体器件。 The apparatus includes an illumination system, the illumination system Zen bonded to the microfluidic device and the electromagnetic emission blessing illuminating the microfluidic device. 该装置还包括成像系统,该成像系统禪合到微流体器件并用于接收从多个处理位置发射过来的电磁福射。 The apparatus further comprises an imaging system, the imaging system Zen and bonded to the microfluidic device for receiving electromagnetic radio-fu transmitted from a plurality of processing positions over. 该装置另外包括禪合到成像系统的检测器。 The apparatus additionally includes a Zen bonded to the imaging system detector.

[0024] 在另一个特定实施例中,提供了一种光学成像系统。 [0024] In another particular embodiment, there is provided an optical imaging system. 该光学成像系统包括计算机和光学照明系统,该光学照明系统用于照射弹性微流体阵列器件,运种器件至少包括1536 个处于流体隔离状态中的反应室。 The optical imaging system includes a computer and an optical illumination system, the optical illumination system for illuminating elastomeric microfluidic array device, such devices are transported in a fluid comprising at least 1,536 of the reaction chamber in isolation. 该弹性微流体阵列器件包括由多个层构成的弹性块。 The elastomeric microfluidic array device comprising an elastomeric block constituted by a plurality of layers. 在运多个层中,至少一层具有形成于其中的至少一个凹陷。 In operation a plurality of layers, at least one layer having formed therein at least one recess. 该凹陷具有至少一个可弯曲的膜, 该膜被整合到凹陷的层上。 The recess having at least one flexible membrane which is integrated into the recessed layer. 光学成像系统还包括光学检测系统。 The optical imaging system further includes an optical detection system.

[00巧]在另一个可选的特定实施例中,提供了一种给微流体器件的多个反应室内的一个或多个选出的巧光指示成像的方法。 [00 Qiao] In another alternative particular embodiment, there is provided a plurality of reaction to one or more microfluidic device chamber elected clever light indicates the method of imaging. 该方法包括:沿成像通道,传输一个或多个巧光发射信号,所述一个或多个巧光发射信号是从微流体器件的多个反应室中的至少一个反应室内的一个或多个样品那儿获得的。 The method comprising: forming along the channel, the transmission of one or more light emitting signal Qiao, Qiao said one or more light emitting signals from a plurality of reaction chambers in the microfluidic device at least one of a plurality of samples of the reaction chamber or get there. 该方法还包括:通过使用滤光器件,沿成像通道,选择性地传输上述一个或多个巧光发射信号的子集,该滤光器件用于使预定光谱带宽内的巧光发射信号通过,并且将与上述一个或多个巧光发射信号相关联的一种或多种色差处理到预定的程度。 The method further includes: using optical devices along the imaging channel, selectively transmitting said one or more light emitting clever subset signal, the optical device for causing a clever light within a predetermined spectral bandwidth of the transmitted signal by, and the said one or one and a plurality of light emitting clever signal associated with one or more color processing to a predetermined level.

[00%] 在特定的实施例中,该方法还包括:在检测器处,读取上述一个或多个巧光发射信号的子集的一部分;拍摄上述一个或多个巧光发射信号,运些信号是从微流体器件的多个反应室中的至少一个反应室内的一个或多个样品那儿获得的;照亮上述至少一个微流体器件中的至少96个反应室,其中在多于48个反应室构成的一组中的每一个反应室与多于48 个反应室构成的另一组中的每一个反应室都处于流体隔离状态,并且使上述至少一个微流体器件在确定的时间维持预定的溫度,其中预定时间处的预定溫度是多步热循环分布中的一部分。 [00%] In a particular embodiment, the method further comprising: at the detector, or to read a portion of the subset of the plurality of light emitting coincidence signal; shoot the one or more light emitting signal clever, transport these signals from the plurality of reaction chambers in the microfluidic device of at least one reaction chamber or a plurality of samples where obtained; illuminating said at least one microfluidic device of at least the reaction chamber 96, in which more than 48 each of a group consisting of the reaction chamber and the reaction chamber 48 other than the group consisting of the reaction chamber of each reaction chamber are in fluid isolation, and the above-mentioned at least one microfluidic device is maintained at a predetermined time determined temperature, wherein the predetermined temperature is a predetermined time at a multi-step thermal cycling profile part. 在另一个特定的实施例中,处理与上述一个或多个巧光发射信号相关联的一种或多种色差的过程包括:使上述一种或多种色差减小到预定的程度。 In another particular embodiment, the process with one or more of one or more light emitting clever color difference signals associated comprising: one or more of the above-mentioned color reduced to a predetermined level. 该预定的程度W预定的焦点移动为特征,而该预定的焦点移动又与具有第一颜色的第一光线和具有第二颜色的第二光线相关联。 The extent to which predetermined W predetermined movement is characterized by the focus, and the focus movement and with a predetermined first light having a first color and a second light having a second color is associated. 在另一个特定的实施例中,滤光器件包括多个零屈光度双透镜和多个光谱滤光片。 In another particular embodiment, the filter device includes a plurality of lenses and a plurality of pairs of zero refractive power spectral filter.

[0027] 在本发明的一个实施例中,使用光学成像系统来收集和微流体器件有关的信息。 [0027] In one embodiment of the present invention, an optical imaging system to collect and related information microfluidic device. 在特定的实施例中,成像系统的使用包括一种由不止63个反应室构成的微流体器件。 In a particular embodiment, using an imaging system comprises a reaction chamber 63 than the configuration of the microfluidic device. 在另一个特定的实施例中,成像系统的使用包括一种具有弹性微流体器件的微流体器件。 In another particular embodiment, using an imaging system includes a microfluidic device having an elastomeric microfluidic device. 在可选的实施例中,成像系统的使用包括由不止95个反应室构成的微流体器件。 In an alternative embodiment, the imaging system comprises the use of more than the reaction chamber 95 consisting of a microfluidic device. 在另一个可选的实施例中,成像系统的使用包括一种由不止383个反应室构成的微流体器件。 In another alternative embodiment, using an imaging system comprises a reaction chamber 383 than the configuration of the microfluidic device. 在另一个可选实施例中,成像系统的使用包括一种由不止511个反应室构成的微流体器件。 In another alternative embodiment of the microfluidic device, used by the imaging system comprises a reaction chamber 511 than the embodiment constituted. 在另外的实施例中,成像系统的使用包括一种由2304个或更多个反应室构成的微流体器件。 In a further embodiment, the use of the imaging system 2304 includes one or more reaction chambers constituting the microfluidic device. 在另一个附加实施例中,成像系统的使用包括一种由9216个或更多个反应室构成的微流体器件。 In another additional embodiment, the use of the imaging system 9216 includes one or more reaction chambers constituting the microfluidic device. 在另一个附加实施例中,成像系统的使用包括一种由100000个或更多个反应室构成的微流体器件。 In another additional embodiment, using an imaging system includes one or more of the 100,000 reaction chambers constituting the microfluidic device.

[0028] 根据本发明的另一个实施例,使用光学成像系统来收集和微流体器件有关的信息。 [0028] In accordance with another embodiment of the present invention, an optical imaging system to collect and related information microfluidic device. 光学系统的使用包括一种弹性微流体器件,运种弹性微流体器件适合于在特定的实施例中执行蛋白质结晶化处理。 Using an optical system comprising an elastomeric microfluidic device, the elastic kind of transport microfluidic device adapted to perform protein crystallization process in certain embodiments. 在另一个实施例中,成像系统的使用包括一种弹性微流体器件,运种弹性微流体器件适合于执行有关处理。 In another embodiment, using an imaging system includes an elastomeric microfluidic device, transported species elastic microfluidic device suitable for the implementation of the relevant process. 在另一个实施例中,成像系统的使用包括一种弹性微流体器件,运种弹性微流体器件适合于在选定的溫度或溫度范围中进行各种反应。 In another embodiment, using an imaging system includes an elastomeric microfluidic device, the elastic kind of transport microfluidic device suitable for performing various reactions at a selected temperature or temperature range. 在另一个特定的实施例中,光学成像系统被用于收集和PCR反应有关的信息。 In another particular embodiment, the optical imaging system is used to collect information about and PCR reactions. 在另一个特定的实施例中,成像系统的使用包括一种微流体器件,运种微流体器件包括处于微流体隔离状态中的封闭式反应室。 In another particular embodiment, using an imaging system includes a microfluidic device, the microfluidic device comprises a kind of operation in the microfluidic isolation of enclosed reaction chamber. 在其它实施例中,成像系统的使用包括禪合到该微流体器件的热控制器。 In other embodiments, using an imaging system includes a controller Zen bound to heat the microfluidic device.

[0029] 在可选的实施例中,使用光学成像系统来给微流体器件成像。 [0029] In an alternative embodiment, an optical imaging system for imaging a microfluidic device. 在运种可选实施例中,光学成像系统具有多个双透镜,适合于将色差的值减小到预定的程度。 In alternative embodiments kind of operation, an optical imaging system having a plurality of pairs of lenses, adapted to the value of the color difference is reduced to a predetermined extent. 在一个实施例中,光学成像系统具有大于0. 23的NA。 In one embodiment, the optical imaging system is greater than 0.23 NA. 在另一个实施例中,光学成像系统具有大于或等于0.36的NA。 In another embodiment, the optical imaging system is greater than or equal to 0.36 of NA. 在另一个实施例中,光学成像系统具有大于0.2的NA。 In another embodiment, an optical imaging system having greater than 0.2 NA. 在特定的实施例中,光学成像系统包括:第一光谱滤光片,禪合到第一双透镜;W及第二光谱滤光片,禪合到第二双透镜。 In a particular embodiment, the optical imaging system comprising: a first spectral filter, Zen bonded to the first pair of lenses; W and the second spectral filter, Zen bonded to a second two-lens. 此外,在另外的实施例中,第一双透镜使第一光谱滤光片所透射的第一波长处的色差值减小到第一预定程度,而第二双透镜使第二光谱滤光片所透射的第二波长处的色差值减小到第二预定程度。 Further, in another embodiment, the first pair of lens color difference value of the first spectral filter being transmitted at the first wavelength is decreased to a first predetermined level, and the second pair of the second lens spectral filter color sheet values are transmitted at the second wavelength is reduced to a second predetermined level. 在某些实施例中,第一双透镜和第二双透镜是零屈光度双透镜。 In certain embodiments, the first dual-lens and the second lens is a double-double zero diopter lens. 在其它实施例中,第一光谱滤光片、第一双透镜、第二光谱滤光片和第二双透镜是作为滤光轮来排列的。 In other embodiments, the first spectral filter, a first double-lens, a second lens pair and the second spectral filter is arranged as a filter wheel.

[0030] 在本发明的另一个实施例中,使用巧光成像系统从微流体器件那儿获取多个图像,其中包括一个或多个巧光发射信号。 [0030] In another embodiment of the present invention, the use of clever optical imaging system to obtain a plurality of images from the microfluidic device there, including one or more clever optical transmission signal. 巧光成像系统的使用包括一种具有不止63个反应室的微流体器件,运些反应室都处于流体隔离状态中。 Clever use of light than the imaging system comprises a reaction chamber 63 having a microfluidic device, are transported in these reaction chambers in fluid isolation. 在一个实施例中,巧光成像系统的使用包括一种具有图像微流体器件的微流体器件。 In one embodiment, the optical imaging system using clever comprises a microfluidic device having a microfluidic device of the image. 在另一个实施例中,巧光成像系统的使用包括多个其体积小于IOnl的反应室。 Examples, clever optical imaging system includes a plurality of volume smaller than the reaction chamber IOnl in another embodiment. 在另一个实施例中,巧光成像系统的使用包括一种弹性微流体器件,运种弹性微流体器件具有每平方厘米大于或等于100个反应室的阵列密度。 In another embodiment, clever use of the optical imaging system comprises an elastomeric microfluidic device, the elastic kind of transport device having an array of microfluidic density per square centimeter is greater than or equal to 100 in the reaction chamber. 在可选的实施例中,巧光成像系统包括一种滤光器件,该滤光器件适合于使上述一个或多个巧光发射信号中选定的光谱带宽透射,并且还适合于将与上述一个或多个巧光发射信号相关联的一种或多种色差处理到确定的程度。 In an alternative embodiment, the clever optical imaging system comprises a filter device, the optical device is adapted to cause the one or more light emitting signal Qiao selected spectral bandwidth transmission, and further adapted to the above or one more of a clever light-emitting signals associated with one or more color processing to determine the extent of.

[0031] 在本发明的一个实施例中,提供了一种对微流体系统中的样品进行询问的光学系统。 [0031] In one embodiment of the present invention, there is provided a microfluidic system samples were asking an optical system. 在本实施例所提供的光学系统中,检测器由一个阵列(例如,CCD阵列)构成,其尺寸等于或大于要被成像到其上的微流体系统。 In the present embodiment the optical system is provided in the case, it consists of a detector array (e.g., the CCD array) form, its size is equal to or greater than the microfluidic system to be imaged thereon. 在另一个实施例中,检测器通过使用至少一个透镜,W光学方式禪合到微流体器件。 In another embodiment, the detector by using at least one lens, W is optically bonded to the microfluidic device Zen. 在另一个实施例中,检测器并不与微流体器件保形接触。 In another embodiment, the detector is not in contact with the conformal microfluidic device. 在特定的实施例中,检测器与微流体器件保形接触。 In a particular embodiment, the detector and the microfluidic device in conformal contact. 在另一个特定的实施例中,在光学系统中使用了在整个说明书中都讨论到的色差校正系统。 In another particular embodiment, the optical system is used throughout the specification are discussed chromatic aberration correction system.

[0032] 在另一个实施例中,提供了一种对微流体器件进行询问的光学系统。 [0032] In another embodiment, there is provided a microfluidic device for questioning optical system. 光学系统具有放大率M > NAyNAdet,其中NA。 An optical system having a magnification M> NAyNAdet, wherein NA. 是物方/样品那一侧的NA,而NA det是在因检测器表面反射所造成的损耗超过阔值之前可允许到达该检测器的最大的NA。 NA is the object-side / side of the sample, while NA NA det is the largest of the detector before the detector surface reflection due to the loss caused by the threshold value exceeds the allowable arrival. 在特定的实施例中,检测器是CCD,并且NAdet= 0. 36。 In a particular embodiment, the detector is a CCD, and NAdet = 0. 36. 在另一个实施例中,检测器是CCD,并且0. 23《NA det《0. 5。 In another embodiment, the detector is a CCD, and 0. 23 "NA det" 0. 5.

[0033] 在附加的实施例中,提供了一种对微流体系统中的样品进行询问的光学系统。 [0033] In an additional embodiment, there is provided a microfluidic system samples were asking an optical system. 检测器包括一种阵列(例如,CCD阵列),其大小等于包含多个反应室的微流体器件的面积的M倍。 It comprises a detector array (e.g., the CCD array), which is equal to the size of the reaction chamber comprising a plurality of microfluidic device area M times. 在一个实施例中,微流体器件的面积包括多个要被询问的样品室。 In one embodiment, the microfluidic device comprises a plurality of areas of the sample chamber to be interrogated. 在该实施例中,M =NAyNAdet+/-l〇%,其中NA。 In this embodiment, M = NAyNAdet +/- l〇%, wherein NA. 是由焦涂等种种考虑所确定的可接雙的最大NA,而焦涂等种种考虑则设及反应室之间可接受的串扰、它们的XY间隔和尺寸W及它们沿Z轴的宽度,并且NAd。 Is connected by a double-maximum NA coke coating and other considerations determined, and the coke coating and other considerations are acceptable provided between the reaction chamber and crosstalk, their XY spacing and size of the W and Z axis along their width, and nAd. ,使得《40%的入射光因反射、晕映等原因而在检测器表面处损失了。 So that "40% of the incident light due to reflection, vignettes and other reasons, the detector surface loss. 在另一个特定的实施例中,NAd。 In another particular embodiment, NAd. ,使得《20%的入射光因反射、晕映等原因而在检测器表面处损失了。 So that "20% of the incident light due to reflection, vignettes and other reasons, the detector surface loss. 在另一个特定的实施例中,NAd。 In another particular embodiment, NAd. ,使得《10%的入射光因反射、晕映等原因而在检测器表面处损失了。 So that "10% of the incident light due to reflection, vignettes and other reasons, the detector surface loss. 在可选的实施例中,M >1。 In an alternative embodiment, M> 1. 在另一个可选的实施例中,1=1。 In another alternative embodiment, a = 1.

[0034] 和常规技术相比,通过使用本发明,实现了大量的益处。 [0034] and compared to conventional techniques, through the use of the present invention, to achieve a number of benefits. 一些实施例提供了光学成像系统,W产生并检测来自微流体器件的巧光。 Some embodiments provide an optical imaging system, W generated and detected light from the coincidence of the microfluidic device. 另外,本发明的各实施例提供了一些光学系统,运些光学系统通过使用禪合到光谱滤光片的零屈光度消色差双透镜来减小色差。 Further, various embodiments of the present invention provides an optical system, the optical system through the use of some operation Zen spectral filter laminated to zero diopter achromatic doublet lens to reduce chromatic aberration.

[0035] 本发明的各实施例能够在单个平台上执行许多应用。 Embodiments [0035] The present invention is capable of performing many applications on a single platform. 例如,通过使用本发明的各实施例,来执行各种基因组应用,其中包括基因表达和基因型鉴定。 For example, through the use of various embodiments of the present invention is to perform a variety of genomic applications, including gene expression and genotyping. 此外,数字隔离与检测值ID)应用是可行的。 Furthermore, the digital isolation and detection value ID) is feasible. 作为一个示例,提供了许多设及癌症检测、单细胞大分子检测和量化等应用。 As an example, it offers a number of set and cancer detection, detection and quantification of cellular macromolecules single applications. 此外,通过本发明的各实施例,提供了许多蛋白质组学的应用,其中包括蛋白质配体结合W及免疫测定处理。 In addition, various embodiments of the present invention, there is provided a number of proteomics applications, including protein and ligand binding W immunoassay process.

[0036] 另外,根据本发明的各实施例,可W实现某些益处和/或优点,W克服其它技术的某些局限。 [0036] Further, according to the embodiment of the present invention, at the W realize certain benefits and / or advantages, W overcome some of the limitations of other techniques. 下面的内容都是基于计算而确定的: The following content is determined based on the calculation:

[0037] 一些扫描系统具有期望的分辨率10微米。 [0037] Some scanning systems having a desired resolution of 10 microns. 在运些系统中,总共有3000x3000(= 约IO67)个"点",期望针对运些"点"来使巧光量化。 In the operation of these systems, a total of 3000x3000 (= about IO67) a "point", it is desirable for the operation of these "points" to make clever light quantization. 例如,为了在10秒内实现上述运一点, 每一个点处的停留时间是~I微秒。 For example, in order to achieve the above-described operation point within 10 seconds, the residence time at each point is ~ I microseconds. 相应地,在3毫秒内将完成线扫描(3000个点)。 Accordingly, within 3 milliseconds to complete the scan line (3000 points). 为了获得和使用本发明各实施例而实现的信噪比等价的信噪比,该"点"处的光强应该比本发明的成像系统通过在1秒周期内获得一个信号从而实现的光强要亮~1〇6倍。 SNR SNR equivalent in order to obtain and use the invention and various embodiments realized, the light that "point" should be stronger than at the imaging system of the present invention by obtaining a signal within 1 second cycle in order to achieve light strong brighter ~ 1〇6 times. 为了在3毫秒内越过3厘米扫描一个点,1000次/秒在机械方面常常是很有挑战性的。 In order to scan across a point 3 cm 3 msec, 1000 times / second is mechanically often very challenging. 另外,期望高效地收集发射信号。 Further, it is desirable to efficiently collect the transmitted signal. 运通常需要很大的场光学镜片或NA很高的浮动头(例如,W>= lOm/s的速度移动)。 Transport usually takes a lot of high-field optical lenses or NA floating head (for example, W> = lOm / s speed of movement). 在运种情况下,和器件左上角处收集的数据相比,器件右下角处收集的数据将晚10秒,运在某些应用中是不期望出现的。 In the transport case, the upper left corner of the device and the data collected as compared to the data collected at the bottom right corner of the device 10 seconds later, shipped in certain applications is undesirable arise.

[0038] 一些缝合系统获取多个图像,并且将它们缝合到一起从而形成复合图像。 [0038] Some suturing system to obtain a plurality of images and stitching them together to form a composite image. 在一个系统中,为了用多个3mmx3mm的图像缝合出一个图像,利用了100个图像。 In a system, for a plurality of image stitching 3mmx3mm an image using the image 100. 获取运些单张图像所用的总时间将是100毫秒。 The total time to acquire some run single image will be used for 100 milliseconds. 假设大约一半的时间用在移动上面,另一半时间用在获取图像上面,则图像时间将大约是50毫秒。 Assumptions used in about half the time to move on top of the other half to the acquisition of the above image, the image of time would be approximately 50 milliseconds. 该系统通常需要比本发明各实施例在1秒周期内获取1个信号的过程中所用的光强要强20倍的光强,和/或还需要NA更高的物镜。 The system typically requires 20 times stronger intensity of the light intensity in Example obtain a signal within one second cycle used in the process and / or need NA higher than the objective lens of the present invention embodiments. 另外, 运些系统通常需要快速的镜台-能够移动3毫米并且能够在50毫秒内完全结束。 In addition, these systems often require rapid transport of the stage - can be moved 3 mm and can be completely finished within 50 milliseconds. 另外,运些系统通常需要对移动和光强进行仔细的校准,W消除缝合过程中的伪像。 In addition, these systems generally require transport to move carefully and intensity calibration, W during suturing eliminate artifacts. 和左上角处收集的数据相比,右下角处收集的数据将晚10秒,运在某些应用中是不期望出现的。 And data collected at the upper left corner compared to data collected at the bottom right corner 10 seconds later, shipped in certain applications is undesirable arise.

[0039] 本发明的一些实施例提供了能够使微流体器件所在区域一次全部成像的成像系统。 Some embodiments [0039] The present invention provides a microfluidic device capable of making the region where the imaging imaging system all at once. 在每个图像可W有10秒的一个实施例中,可W有若干秒的整合时间。 W in each image 10 of a second embodiment, at the W of a number of second integration time. 同步化问题减小了或消除了,因为数据是同时从所有的像素中收集的。 Synchronization problem is reduced or eliminated, because the data is collected simultaneously from all the pixels. 此外,本发明的各实施例通常同时照亮整个微流体器件(~31mmx31mm的区域),并且使用具有高NA和大视场的透镜系统。 In addition, embodiments of the present invention generally simultaneously illuminate the entire microfluidic device (~ 31mmx31mm region), and having a high NA and a large field of view of the lens system. 如本说明书中所描述的,本发明的各实施例提供了比常规系统简单且更有用的方法和系统。 As described in this specification, embodiments of the present invention provide a simpler and more useful than conventional systems approach and systems. 根据各实施例,本发明各实施例的方法和系统可W克服上述运些局限中的一种或多种。 Methods and systems according to various embodiments, various embodiments of the invention can overcome the above W transported some limitations in one or more. 在本说明书中,尤其是在下文中,可W找到本发明的细节。 In the present specification, especially in the following, W can find details of the invention. 根据各实施例,可能存在运些益处中的一种或多种。 According to various embodiments, there may be some benefits in the operation of one or more. 在本说明书中,尤其是在下文中,会描述运些和其它益处。 In the present specification, especially in the following, the operation will be described and some other benefits.

[0040] 参照下面的详细描述和附图,可W更全面地理解本发明的各种其它的目的、特征和优点。 [0040] reference to the following detailed description and drawings, W can be more fully understood the invention for various other objects, features and advantages.

附图说明 BRIEF DESCRIPTION

[0041] 图IA是示出了本发明一实施例的光学成像系统的简化示意图; [0041] FIG. IA is a simplified schematic diagram illustrating an optical imaging system according to an embodiment of the present invention;

[0042] 图IB描绘了本发明一可选实施例的典型成像系统的概观; [0042] FIG. IB depicts the present invention, an alternative exemplary embodiment of the imaging system overview;

[0043] 图IC是示出了本发明一实施例的热控制器件的简化示意图; [0043] FIG. IC is a simplified schematic diagram illustrating a thermal control device according to an embodiment of the present invention;

[0044] 图2A-2C是示出了本发明一实施例的透镜组件的简化示意图; [0044] FIG. 2A-2C is a simplified schematic diagram illustrating a lens assembly according to an embodiment of the present invention;

[0045] 图3是根据本发明一实施例在微流体器件的许多反应室中通过反应而产生的W 第一波长为中屯、的巧光发射的照片; [0045] FIG. 3 is a first wavelength W in many cases the reaction chamber microfluidic devices produced by reaction of one embodiment of the present invention is in the village, clever light emission photograph;

[0046] 图4是根据本发明一实施例在微流体器件的许多反应室中通过反应而产生的W 第二波长为中屯、的巧光发射的照片; [0046] FIG. 4 is a second embodiment of the present invention W wavelength in many cases the reaction chamber microfluidic devices produced by the reaction of the village, the light-emitting clever photos;

[0047] 图5-7是使用本发明一实施例而产生的选定波长的光点图; [0047] FIG. 5-7 is selected wavelengths of light dot pattern using an embodiment of the present invention produced;

[0048] 图8是使用本发明一实施例而产生的照明图,该照明图示出了相对均匀性和位置的函数关系图; W例图9-11是本发明若干实施例的正方块块内能量图; [0048] FIG. 8 is an embodiment of the present invention generated illumination pattern, the illumination diagram illustrating a function of the relative homogeneity and location; W is the number of cases of 9-11 square tranches embodiment of the present invention The energy diagram;

[0050] 图12是示出了本发明一实施例所提供的光学系统的场曲和崎变的图; [0050] FIG. 12 is a diagram showing the present invention, curvature of field and Kawasaki varying view of an optical system provided in an embodiment;

[0051] 图13是示出了本发明一实施例的系统所产生的"双波长对焦点"的图; [0051] FIG. 13 is a diagram showing a system according to the present invention produced by the "dual-wavelength focus" according to one example;

[0052] 图14-16是使用本发明一实施例而产生的选定波长的光点图; [0052] Figures 14-16 is a selected wavelength of light spot diagram of the invention using embodiments of generated;

[0053] 图17-19是使用本发明一实施例而产生的选定波长的光点图; [0053] Figures 17-19 is a selected wavelength of light spot diagram of the invention using embodiments of generated;

[0054] 图20-22是使用本发明一实施例而产生的选定波长的光点图; [0054] 20-22 is selected wavelength light spot diagram of the invention using embodiments of generated;

[0055] 图23-25是使用本发明一实施例而产生的选定波长的光点图; [0055] Figures 23-25 is a selected wavelength of light spot diagram of the invention using embodiments of generated;

[0056] 图26-28是本发明若干实施例的正方块块内能量图。 [0056] The present invention is 26-28 square several tranches within the energy diagram of an embodiment.

具体实施方式 detailed description

[0057] 根据本发明,提供了用于微流体系统的多种技术。 [0057] According to the present invention, there is provided a variety of techniques for microfluidic systems. 特别是,本发明提供了一种给微流体器件的反应室内的流体体积中悬浮的一个或多个实体成像的方法和系统。 In particular, the present invention provides a method and system to the fluid volume of the reaction chamber of the microfluidic device suspended one or more entities imaging. 更具体地讲,运种成像方法和系统利用了与微流体器件中的一个或多个实体相关联的巧光信号的指示。 More specifically, the image forming method and system of transport use microfluidic device indicating that one or more entities clever optical signal associated. 仅仅作为示例,通过使用和微流体器件相禪合的巧光、化学发光和生物发光读出器,来应用微流体方法和系统技术,但是将会认识到本发明具有更广的应用范围。 Just as an example, by using a microfluidic device and meditation combined with clever light, chemiluminescence and bioluminescence reader to apply microfluidic methods and systems technology, it will be recognized that the invention has a wider range of applications.

[005引在本申请中,提到了微流体器件中的某些类型的"反应"室。 [005 incorporated in the present application, reference is made microfluidic device of some type of "response" room. 通常,运些"反应室" 包括处理位置、处理室、和/或反应位置、它们的任意组合等等。 Typically, some op "reaction chamber" includes a processing position, the process chamber, and / or reaction site, any combination thereof and the like. 运些室室可W是闭合的、部分闭合的、打开的、部分打开的、密封的、或它们的组合等等,其中包括设及运些状态中的任何状态的任何暂时或瞬时条件。 These rooms can transport chamber W is closed, partially closed, open, partially open, closed, or a combination thereof, and so on, including the set up and operation of any of these conditions are temporary or transient state of any state. 在一些实施例中,运些室室是密封的、能够密封的、可闭合的、隔离的、能够隔离的、和它们的组合、W及设及运些状态中的任何状态的任何暂时或瞬时条件中的任何组合或单个条件。 In some embodiments, transported some room chamber is sealed, can seal can be closed, isolated, can be isolated, and combinations thereof, W, and set up and run any state some state of any temporary or transient conditions in any combination or single condition. 因此,术语"反应室"的使用并不旨在限制本发明,而是包括运些其它结构。 Thus, the term "reaction chamber" is not intended to limit the use of the present invention, but include some other transport structure. 另外,术语"室"并不旨在限制本发明,而是应该按其普通的意思来使用,除非已经引用和该室相关联的特定特征。 In addition, the term "room" is not intended to limit the invention, but should be used according to their ordinary meaning, unless specific features and references associated with the chamber. 当然,可W有其它变化、修改和替换。 Of course, there are other W changes, modifications and substitutions.

[0059] 此外,在本申请中,提到了来自微流体器件的巧光指示。 [0059] In the present application, it refers to an instruction from the clever optical microfluidic devices. 本发明的范围内不仅包括巧光指示,还包括发光指示,其中包括化学发光、电致发光、电化学发光、憐发光、生物发光和其它发光过程、或包括可用检测器件检测的任何其它类型的指示的任何其它过程。 Included within the scope of the present invention is not only clever and light indication, including flares, including chemiluminescence, electroluminescence, ECL, pity, bioluminescence and other emission process, or include any other type of detection device detects the available any other process indication. 对于本领域的技术人员,很明显,可用于检测并分析运些巧光和发光指示的方法和系统可W从一种指示转移到另一种指示。 For those skilled in the art, obviously, it can be used to detect and analyze these transport methods and systems can be clever light emitting directions W transfer from one indication to another indication. 另外,尽管本发明的一些实施例利用光谱滤光片作为光学元件,但是本发明并没有要求运一点。 In addition, although some embodiments of the present invention utilizes an optical spectral filter element, but the present invention does not require transport point. 一些巧光和发光应用并不在光激发通道、光发射通道或二者中使用光谱滤光片。 Some applications are not clever light-emitting light excitation channels, or both channel light emission spectral filter is used. 如本文所述,其它实施方式采用光谱滤光片。 As described herein, other embodiments using spectral filter. 本领域的技术人员会理解与特定应用相关联的诸多差异。 Those skilled in the art will appreciate that many differences associated with specific applications.

[0060] 在一些实施方式中,使用了用于进行微流体分析的各种器件和方法,其中包括可用于进行热循环反应如核酸扩增反应的器件。 [0060] In some embodiments, using a variety of devices and methods for microfluidic analysis can be used, including thermal cycling reactions such as nucleic acid amplification reaction device. 运些器件和常规微流体器件相比不同之处在于:它们包括了弹性组件;在某些情况下,该器件大部分或全部都是由弹性材料构成的。 These transport device compared to conventional microfluidic device except that: they include elastomeric components; in some cases, most or all of the device is made of an elastic material. 例如,扩增反应可W是线性扩增(用一种引物扩增)W及指数扩增(即用正向和反向组进行扩增)。 For example, the amplification reaction may be W is a linear amplification (using a primer) W and exponential amplification (ie, groups with forward and reverse amplification).

[0061] 本发明的一些实施方式所提供的方法和系统在执行核酸扩增反应的过程中利用了盲端通道类型的器件。 [0061] The method and system of some embodiments of the present invention, there is provided the use of the blind end of the channel type of device during the execution of a nucleic acid amplification reaction. 在运些器件中,通常沉积在反应位点内的试剂是进行所需类型的扩增反应所必需的那些试剂。 In operation these devices usually deposited reagents in the reaction sites are those reagents required for the type of amplification reaction was necessary. 通常,运意味着沉积W下某些或全部试剂:引物、聚合酶、核巧酸、金属离子、缓冲液和辅因子。 Typically, the transport means that some or all of the reagents under the deposition W: primer, polymerase, nuclear clever acids, metal ions, buffers and cofactors. 在运种情况下,引入反应位点的样品是核酸模板。 In the transport case, the sample is introduced reactive sites is a nucleic acid template. 然而,可沉积该模板,并且使扩增试剂流入反应位点。 However, it may be deposited on the template, and the amplification reagents into the reaction sites. 如本发明中详细讨论的那样,当使用矩阵器件进行扩增反应时,含核酸模板的样品流过垂直的流动通道,而扩增试剂则流过水平的流动通道,反之亦然。 As discussed in detail in the present invention, when using a matrix devices amplification reaction, a sample containing nucleic acid template flowing perpendicular to the flow channel, and the amplification reagent flows through horizontal flow channel, and vice versa.

[0062] PCR或许是已知最好的扩增技术。 [0062] PCR is perhaps the best known amplification techniques. 本发明各实施例所使用的器件并不限于进行PCR扩增。 Device used in Example embodiments of the present invention is not limited to PCR amplification. 可W进行的其它类型的扩增反应包括但不限于:(i)连接酶链反应化CR)(参照Wu 和Wallace, Genomics 4:560(1989) W 及Landegren 等人,Science 241:1077(1988)); (ii)转录扩增(参照Kwoh 等人,Proc. Natl. Acad. Sci. USA 86:1173(1989)) 自动维持序列复制(参照Guatelli 等人,Proc.化t. Acad. Sci. USA, 87 :1874(1990)) ; W及(iv)基于核酸的序列扩增(NASBA)(参照Sooknanan,R.和Malek,L.,BioTechnology 13 : 563-65 (1995))。 Other types of amplification reactions can be carried W include but are not limited to: (i) ligase chain reaction of the CR) (refer to Wu and Wallace, Genomics 4: 560 (1989) W and Landegren et al, Science 241: 1077 (1988 )); (II) transcription amplification (see Kwoh et al., Proc Natl Acad Sci USA 86: 1173 (1989)) maintained automatically sequence replication (see Guatelli et al., Proc of t Acad Sci........ USA, 87: 1874 (1990)); W and (iv) a nucleic acid sequence-based amplification (the NASBA) (refer to Sooknanan, R and Malek, L., BioTechnology 13: 563-65 (1995.)). 上述各种参考文献W其全部内容引用在此。 The above-described various references W content reference herein in its entirety.

[0063] 此外,设计某些器件,W采用包括一个或多个弹性阀的器件进行热循环反应(如PCR),所述弹性阀用来调控流过该器件的溶液。 [0063] In addition, the design of certain components, W comprises the use of one or more elastic devices valves thermal cycling reaction (e.g., the PCR), said resilient valve for the regulation of solution flow through the device. 因此,也提供了用运种设计的器件来进行扩增反应的方法。 Thus, also provided with a device designed to transport species amplification reaction.

[0064] 不管是被隔离在反应室内还是在随后的任何时刻,都可W用检测核酸的常规方法来检测并区分扩增产物(扩增子)。 [0064] is isolated in the reaction either indoors or at any subsequent time, the conventional method of detecting a nucleic acid W can be used to detect and distinguish between the amplification product (amplicon). 使用嵌入染料如SYBR™、Pico Green(俄勒冈州Elugene 市Molecular Probes 有限公司)、漠化乙锭等(参照Zhu 等人,1994, Anal. Qiem. 66 : 1941-48)和/或凝胶电泳,可W检测包含双链DNA的扩增子。 Using embedded dyes such as SYBR ™, Pico Green (Molecular Probes, Oregon City Elugene Ltd.), such as ethidium desertification (see Zhu et al., 1994, Anal Qiem 66:.. 1941-48) and / or gel electrophoresis, W detect amplicon may contain double-stranded DNA. 更常见的是,使用序列特异性检测方法(即基于核巧酸序列检测扩增子)。 More common is the use of sequence-specific detection method (ie, nuclear Qiao acid sequence-based detection amplicons). 检测方法的示例包括:固定的寡核巧酸或多核巧酸的阵列杂交;W及使用基于差别标记分子信标的检测系统,或者其它"巧光共振能量转移"(FRET)检测系统。 Examples of detection method comprising: a fixed oligonucleotide or polynuclear Qiao Qiao acid acid array hybridization; W and use the "Transfer clever FRET" (FRET) detection system based on the difference detection system labeled molecular beacons, or other. 根据本发明的一些实施方式,FRET检测是较佳的检测方法。 According to some embodiments of the present invention, FRET detection is the preferred detection method. 在FRET试验中,检测供体/受体巧光团对中供体(报道物)和/或受体(泽光剂)巧光团的巧光变化。 In FRET experiments, testing the donor / acceptor pair clever light group donor (reporter) and / or receptor (Ze Light Stabilizers) Qiao Qiao light group of light changes. 选择供体和受体巧光团,使得供体的发射光谱与受体的激发光谱交迭。 Select the donor and acceptor clever light group, so that the excitation spectrum overlaps the emission spectrum of the donor and acceptor. 因此,当使运对巧光团彼此靠得足够近时,会发生从供体到受体的能量转移,并且可W检测到运种能量转移。 Thus, when the operation of clever optical group close enough to each other occurs when the energy transfer from the donor to the acceptor, and W can be detected species transported energy transfer. 已知各种试验,包括但不限于:模板延伸反应、定量RT-PCR、分子信标(Molecular Beacons)和侵入检测(Invader assay),下面对它们进行简述。 Various tests are known, including, but not limited to: template extension reactions, quantitative RT-PCR, molecular beacon (Molecular Beacons) and intrusion detection (Invader assay), following them briefly.

[0065] FRET和模板延伸反应使用:用供体/受体对中的一个成员标记的引物,W及用此供体/受体对中的另一成员标记的核巧酸。 [0065] FRET and template extension reactions use: with donor / acceptor pair a member of the labeled primer, W and nuclear use this donor / acceptor pair for the other members of the tag clever acid. 在模板依赖性延伸反应期间将标记核巧酸渗入引物中之前,先将供体和受体分隔得足够远,W使能量转移无法发生。 During the template-dependent extension reaction in the nuclear clever tag penetrate acid primer before, the first donor and acceptor are separated far enough, W so that energy transfer can not occur. 然而,如果标记核巧酸渗入引物并且间隔足够近,则会发生能量转移并且可W检测到运种能量转移。 If, however, marked nuclear skillfully infiltrated acid primer and spaced close enough, the energy transfer can occur and can be detected in W op kind of energy transfer. 运些方法在检测单核巧酸多态性的单碱基对延伸反应中特别有用,参见美国专利5, 945, 283和PCT 公开WO 97/22719。 These transport methods in detecting polymorphisms mononuclear clever acid is particularly useful for a single base extension reaction, see U.S. Patent No. 5, 945, 283 and PCT Publication WO 97/22719. 通过使用本说明书中所述的溫度控制方法和装置,可W任选地对运些反应进行热循环,W增大信号。 By using the temperature control method and apparatus described in this specification can optionally be transported W thermal cycling of these reactions, W increases signal.

[0066] 也可W使用各种所谓的"实时扩增"方法或"实时定量PCR"方法,通过测量扩增过程期间或之后所形成的扩增产物的量,来确定样品中存在的祀核酸量。 [0066] W may also be using a variety of so-called "real time amplification" methods or "real-time quantitative PCR" method, the amount of amplification product formed during or after the process by measuring the amplified nucleic acid in the sample to determine the presence of worship the amount. 发巧光的核酸酶试验是实时定量方法的一个特定示例,实时定量方法可W和本文所述器件一起成功地使用。 Nuclease clever light hair test is a specific example of a method of quantitative real-time, real-time quantitative W method and the device described herein together with success. 运种监控扩增产物的形成的方法包括:用双标记的巧光寡核巧酸探针连续测量PCR产物累积,此方法在文献中常被称为"化qMan "法。 The method of monitoring the formation of amplification products shipped species include: the art of double labeled oligonucleotide light nuclei acid probe clever continuous measurement of PCR product accumulation, this method is often called in the literature "of qMan" method. 参见例如,美国专利5, 723, 591。 See, e.g., U.S. Patent No. 5, 723, 591.

[0067] 在使用分子信标的情况下,当它杂交到扩增产物的互补区域时探针构象的变化产生了可检测的信号。 [0067] In the case of using molecular beacons, when it is hybridized to the amplification product complementary regions probe conformational changes produce a detectable signal. 探针自身包括两部分:5'端的一部分,W及3'端的另一部分。 Probe itself includes two parts: the 5 'portion, W, and ends 3' end of the other part. 运些部分侧接于退火到探针结合位点的探针部分,并且彼此互补。 Transport those portions flanking portion of the probe annealing to the probe binding site, and complement each other. 一端部分通常连接于报道染料, 而另一端部分通常连接到泽光染料。 One end portion is generally connected to a reporter dye and the other end portion is typically connected to the Chak light dye. 在溶液中,运两个端部分可W彼此杂交形成发夹环。 In solution, the two end portions can be transported W hybridized to each other form a hairpin loop. 在运种构象中,报道物和泽光剂靠得足够近,使得报道物的巧光被泽光染料有效地泽灭。 In operation conformers, the reporter and light stabilizers Chak close enough so that the coincidence of light was reported to be effective in Chak Chak off light dye. 相反,杂交探针产生线性化构象,其中泽灭程度降低。 In contrast, hybridization probes produce linearized conformation, which reduces the degree of Chak off. 由此,通过监测运两种染料的发射光变化,有可能间接地监测扩增产物的形成。 Thus, by monitoring changes in the light emission operation of the two dyes, it is possible to indirectly monitor the formation of amplification product. 运种类型的探针及其使用方法在下面的文献中有进一步的描述:如Piatek 等人,1998 年,化t. Biotechnol. 16 :359-63 ;Tyagi 和Kramer, 1996 年,化t. Biotechnology 14 :303-308 ; W及Tyagi 等人,1998 年,化t. Biotechnol. 16 : 49-53(1998)〇 Yun types of probes and their use in the following method is further described in the literature: eg Piatek et al., 1998, of t Biotechnol 16: 359-63; Tyagi and Kramer, 1996 years of t Biotechnology... 14:. 303-308; W and Tyagi et al., 1998, of t Biotechnol 16: 49-53 (1998) square.

[0068] 下列文献中描述了Sco巧ion检测法:Thelwell等人,2000年,Nucleic AcidsResearch,28 :3752-3761 ; W 及Solinas 等人,2001 年,"SNP 分析和FRET 应用中的双链体Sco巧ion 引物"值uplex Sco巧ion primers in SNP analysis and FRETa卵lications),Nucleic Acids Research 29:20。 [0068] The following described in the literature Sco clever ion detection method: Thelwell et al., 2000, Nucleic AcidsResearch, 28: 3752-3761; W and Solinas et al., 2001, "SNP analysis and FRET applications duplexes Sco clever ion primer "value uplex Sco clever ion primers in SNP analysis and FRETa eggs lications), Nucleic Acids Research 29:20. Sco巧ion 引物是巧光PCR 引物,其中探针元件通过PCR终止物连接到5'端。 Sco Qiao Qiao light ion primer PCR primers, wherein the probe element is connected to the 5 'end by PCR Terminator. 它们用于均一溶液中PCR产物的实时扩增子特异性检测。 They are used in a homogeneous solution of real-time PCR amplicon product specifically detected. 可能有两种不同的形式,即"茎-环"形式和"双链体"形式。 There may be two different forms, namely "stem - loop" forms and "duplex" form. 在运两种情况下, 探测机制都是作用于分子内。 In both cases the operation, detection mechanisms are applied to the molecule. 所有形式的Scorpion基本元件是:(i)PCR引物;(ii)PCR终止物,用于防止探针元件的PCR通读(read-t虹OU曲);(iii)特定探针序列;W及(iv)巧光检测系统,该系统包含至少一个巧光团和泽光剂。 Scorpion is the basic element of all forms of: (i) PCR primers; (II) PCR was terminated, to prevent PCR read (read-t Rainbow OU song) probe element; (iii) a specific probe sequence; and the W ( iv) clever optical detection system, the system comprising at least one light group and clever Ze light stabilizers. 在Sco巧ion引物的PCR延伸之后,所得扩增子包含与探针互补的序列,在各PCR循环的变性阶段将其分解成单链。 After PCR Sco clever ion primer extension, the resulting amplified sequence contains a promoter probe complementary denaturation step in each PCR cycle will be broken down into single strands. 在冷却时,该探针可自由地结合该互补序列,从而增加巧光,因为泽光剂不再位于巧光团附近了。 Upon cooling, the probe can be freely combined with the complementary sequences, thereby increasing the clever light, because light Chak agent is no longer in the vicinity of the light group coincidence. PCR终止物防止化q DNA聚合酶进行不期望的探针通读。 PCR terminator of q DNA polymerase prevents undesirable read probe. W例侵入检测(Third Wave Technologies公司,Madison市,威斯康星州)被特别地用于SNP基因型分析,它利用与祀核酸值NA或RNA)或多态性位点互补的寡核巧酸,称为信号探针。 W Example intrusion detection (Third Wave Technologies company, Madison city, Wisconsin) is specifically for SNP genotyping, which uses the value NA worship nucleic acid or RNA) or polymorphism loci complementary oligonucleotide Qiao acid, said a signal probe. 第二寡核巧酸(称为侵入寡核巧酸)包含相同的5'核巧酸序列,但3'核巧酸序列包含核巧酸多态性(位点)。 The second oligonucleotide Qiao acid (called invasive oligonucleotide Qiao acid) contain the same 5 'clever nuclear acid sequence, but the 3' nuclear Qiao Qiao nuclear acid sequence comprising acid polymorphisms (sites). 侵入核巧酸干扰信号探针与祀核酸的结合,使得信号探针的5'端在包含多态性(位点)的核巧酸处形成"下垂物"。 Invasive nuclear clever combination of acid probe signal interfering with worship nucleic acid, so that the signal probe 5 'end to form a "sag was" containing polymorphisms (sites) at the nuclear clever acid. 该复合物被一种称为Cleavase 酶的结构特异性内切核酸酶识别。 Endonuclease recognition specificity within the composite structure is called Cleavase enzyme. Cleavase酶切下核巧酸的5'下垂物。 Under Cleavase acid digestion nuclear clever 5 'was sagging. 所释放的下垂物与携带FRET标签的第S探针结合,由此形成另一种由Cleavase酶识别的双链体结构。 Sag was released and the label carrying the first FRET probe S, thereby forming another duplex structure recognized by the Cleavase enzyme. 此时,Cleavase酶从泽光剂中切下巧光团,产生巧光信号。 At this time, Cleavase enzyme cut clever light Ze group from light agent to produce clever optical signal. 对于SNP基因型分析,设计信号探针,使其与参比(野生型)等位基因或变异(突变)的等位基因杂交。 For SNP genotyping, the signal probe is designed so that the reference (wild type) allele or the variant (mutant) allele hybrids. 不像PCR,有一种线性的信号扩增,其中没有核酸扩增。 Unlike the PCR, there is a linear amplification of signal, in which no nucleic acid amplification. 足W指导本领域普通技术人员的诸多细节参见:Neri, BF*.等人,Advances in Nucleic Acid and Protein Analysis3826 :117-125,2000 年;W 及美国专利6, 706, 471。 Foot W guide many details to those of ordinary skill in See: Wait Neri, BF * people, Advances in Nucleic Acid and Protein Analysis3826: 117-125,2000 years; W and U.S. Patent No. 6, 706, 471.

[0070] 各种多重扩增系统可与本发明联用。 [0070] various multiplex amplification system associated with the present invention may be used. 在一种类型中,可使用多个标记不同的探针(每一个探针被设计成只杂交到特定的目标)同时检测若干个不同的祀标。 In one type, you can use a plurality of different labeled probes (Each probe is designed to hybridize only to a specific target) simultaneous detection of several different worship mark. 因为各探针具有不同的标记,所W根据巧光信号检测与各祀标的结合。 Because each probe has a different label, the W clever combination according to the optical signal detection and worship each subject. 通过明智地选择所使用的不同标记,可进行在单次反应中W不同波长激发和/或检测不同标记的各种分析。 By judicious choice of different markers used can be carried out in a single reaction W different wavelength of the excitation and / or detection of different markers of various analyzes. 例如,可W参照:《巧光光谱学》(FluorescenceSpectroscopy) (Pesce 等人著)Ma;rcel Dekker, New 化rk,(1971) ;White等人著,《巧光分析:实践方法》(Fluorescence Analysis :A Practical Approach) ,MarcelDekker, New York, (1970) !Berlman 著,《芳族分子的巧光光谱手册》 (Handbookof Fluorescence Spectra of Aromatic Molecules),第二片反,Academic Press, 化wYork,(1971) !Griffiths 著,《有机分子的颜色和构造》(Colour and Consti1:utionof Organic Molecules) ,Academic Press,New York, (1976);《指不物》(Indicators) (Bishop 编),化rgamon Press, Oxford, 19723 ; W及Haugland著,《巧光探针和研究用化学物质手册》(Handbook of Fluorescent Probes and ResearchChemicals), Molecular Probes, Elugene 市(1992)。 For example, W reference: "a clever optical spectroscopy" (FluorescenceSpectroscopy) (Pesce et al.) Ma; rcel Dekker, New technology rk, (1971); White et al., "A clever optical analysis: practices" (Fluorescence Analysis :! a Practical Approach), MarcelDekker, New York, (1970) Berlman the "aromatic molecules clever light spectrum Handbook" (Handbookof Fluorescence spectra of aromatic molecules), a second sheet anti, Academic Press, of wYork, (1971 !) Griffiths of "organic molecules color and structure" (Colour and Consti1: utionof organic molecules), Academic Press, New York, (1976); "means do not matter" (Indicators) (Bishop Ed.), of rgamon Press, oxford, 19723; W and Haugland forward, "a clever optical probe and research with chemicals Handbook" (Handbook of Fluorescent probes and ResearchChemicals), Molecular probes, Elugene City (1992).

[0071] 许多与基因组修饰有关的疾病,无论是宿主有机体的抑或感染有机体的疾病,是少量核巧酸发生变化(常常包括单个核巧酸改变)的结果。 [0071] Many diseases related genomic modification, whether it is the host organism or whether the infection disease organism is the result of a small amount of nuclear clever acid change (often including mononuclear Qiao acid change) of. 运种单核巧酸变化称为单核巧酸多态性(简称为SNP),并且发生SNP的位点通常称为多态性位点。 Qiao Yun species mononuclear acid changes called single nuclear polymorphisms Qiao acid (abbreviated as SNP), and the occurrence of the SNP loci commonly referred to as polymorphic sites. 本文所述的器件可用于确定运种多态性位点处存在的核巧酸的身份。 Device described herein can be used to determine the transportation of nuclear species Qiao acid identity exists at polymorphic sites. 作为运种能力的扩展,运些器件可用于基因型分析。 As an extension of the ability to transport species, these devices can be used to transport genotype analysis. 基因型分析包括:确定二倍体生物体(即各基因有两个拷贝的生物体)是否包含两个拷贝的参比等位基因(参比型纯合子),或者包含参比和变异等位基因各一个拷贝(即杂合子),抑或包含两个拷贝的变异等位基因(即变异型纯合子)。 Genotype analysis includes: determining a diploid organism (ie two copies of each gene in the organism) contains two copies of the reference allele (homozygous reference), or the like containing the reference and variable sites one copy of each gene (ie, heterozygous), or contains two copies of the mutant alleles (ie homozygous variant). 当进行基因型分析时,可用本发明方法询问单个变异位点。 When genotype analysis, the method of the present invention can be used to ask a single mutation site. 然而,如下文关于多道传输那部分所进一步描述的,运些方法也可用于确定许多不同的DNA位置(loci)(相同基因上的DNA位置、不同基因上的DNA位置或二者的组合)中单个(基因)的基因型。 However, as discussed below with respect to that part of multi-channel transmission further described, these methods may also be used to transport a number of different DNA to determine the location (loci) (DNA position on the same gene, or a combination of both DNA position on different genes) single (gene) genotypes.

[0072] 用于进行基因型分析的器件被设计成采用大小合适的反应位点,W在统计学上确保该反应位点中存在可行DNA浓度的二倍体的两种等位基因各一拷贝。 A copy of each device [0072] for performing genotype analysis is designed to use the right size of reactive sites, W to ensure the presence of viable DNA concentration of the reaction sites in the diploid statistically two alleles . 否则,分析可能产生表明杂合子就是纯合子的结果,仅仅因为该反应位点处不存在第二种等位基因的拷贝。 Otherwise, the analysis may yield results indicate that heterozygous homozygous, just because of the reaction at the site of the second copy of the allele does not exist. 下表1指出本文所述器件可利用的各种示范性DNA浓度下1纳升反应体积中存在的基因组的拷贝数。 The following table 1 indicates the copy number of the DNA concentration under various exemplary devices described herein can be used in a nanoliter volume of the reaction present in the genome.

[0073] 表格1 :在所示DNA浓度下1纳升体积中存在的基因组拷贝数。 [0073] Table 1: Number of genomic DNA copies at a concentration of 1 nanoliter volumes shown in the present.

[0074] [0074]

Figure CN101793826BD00141

[00巧]一般说来,因样品的随机配比,扩增反应开始之前存在的拷贝数决定了测量中可能的误差。 [00 Qiao] In general, the ratio of the sample by random amplified the number of copies before the start of the reaction determines the measurement of the presence of possible errors. 通常用DNA浓度约为0. 10 yg/ y L的样品来进行使用某些器件的基因型分析, 尽管本发明人已经在每一反应位点有一个基因组的浓度下成功地运行了化qMan反应。 Usually DNA concentration of about 0. 10 yg / y L samples for genotyping using certain devices, although the present inventors have a concentration in the genome of each reaction site next successful run of qMan reaction .

[0076] 基因型分析可W使用各种不同的方法来进行。 [0076] W genotype analysis can use a variety of methods. 在运些方法中,通常足W获得"是" 或"否"的结果,即检测过程只需要能够回答是否存在给定等位基因的问题。 In the operation of these methods usually get enough W "yes" or "no" outcome, namely the detection process only needs to be able to answer whether a given allele problem. 因此,只有使用用来检测可能位于多态性位点处的一种等位基因的存在所必需的引物或核巧酸,才能够进行各种分析。 Therefore, the only possible use for detecting the presence of an allele polymorphism located at the site of the necessary primers or nucleic acid clever to be able to carry out all kinds of analysis. 然而,更常见的是,包括了用于检测多态性位点处可能存在各等位基因的引物和核巧酸。 However, more often, including for detecting polymorphisms may exist for each allele primer and nuclear clever acids.

[0077] 单碱基对延伸(SBP巧反应是一种专口针对基因型分析而开发的技术。尽管已经开发出许多SPBE试验,但是一般方法相当相似。通常,运些试验包括杂交一种与祀核酸互补的引物,使得该引物的3'端恰恰是变异位点的5'端或与其相邻。在一种或多种标记的不可延伸的核巧酸和聚合酶的存在下进行延伸,运种核巧酸与占据变异位点的核巧酸互补。不可延伸的核巧酸是一种核巧酸类似物,一旦将该核巧酸类似物渗入引物中便可W防止聚合酶的进一步延伸。如果所添加的不可延伸的核巧酸与变异位点的核巧酸互补,则标记的不可延伸的核巧酸被渗入该引物的3'端,从而产生标记的延伸产物。因此,延伸引物可W表示哪一种核巧酸存在于祀核酸的变异位点处。运些方法和相关的方法在下列文献中进行了讨论:美国专利5, 846, 710、6, 004, 744、5, 888, 819、5, 856, 092、5, 710, 028 W及WO 92/16657。 [0077] single base pair extension (SBP clever reaction is a special port for genotype analysis and the development of technology. Although many have been developed SPBE test, but the general method is very similar. Typically, these tests include hybridization one kind of transport and worship complementary nucleic acid primer so that the primer 3 'end is precisely the point mutations 5' end or adjacent thereto. in the non-nuclear extending one or more markers clever acids and under the presence of a polymerase extension Yun Qiao acid with nuclear species occupy clever variation site nuclear acid complementary to the nuclear non-extendable Qiao Qiao acid is a nuclear acid analogs, once the nuclear Qiao acid analogs can penetrate primer polymerase W prevent further extend if nuclear inextensible added Qiao acid variable sites of nuclear clever acids complementary to the nuclear non-extension of the mark Qiao acid infiltrated the 3 'end of the primer to produce an extension product mark. Thus, extending primers which W represents a clever nuclear nucleic acids present in worship at the site of the mutation operation of these methods and related methods are discussed in the following documents: U.S. Patent No. 5, 846, 710,6, 004, 744,5 , 888, 819,5, 856, 092,5, 710, 028 W and WO 92/16657.

[0078] 也可用定量PCR方法进行基因型分析。 [0078] The method can also be used quantitative PCR genotyping analysis. 在运种情况下,与各等位基因形式互补的区别标记探针与引物、核巧酸和聚合酶一起被包括成为试剂。 In the transport case, with the difference between the allelic forms of complementary labeled probes and primers, and polymerase nuclear clever acids are included together into a reagent. 然而,可W只用一种探针进行反应,尽管运会产生不确定性,即没有信号是因为没有特定的等位基因还是简单地因为反应失效。 However, the W using only one probe reaction, despite ship create uncertainty that there is no signal because there is no specific alleles or simply fail because of the reaction. 对于一个多态性位点有两个等位基因的典型双等位基因情况,试剂混合物中通常包含两个区别标记的探针(各自与等位基因之一完美互补),W及扩增引物、核巧酸和聚合酶。 For a polymorphic site typically has two alleles of biallelic case, the reagent mixture typically contains two probes diacritical marks (each with one allele perfect complement), W and amplification primers nuclear clever and acid polymerase. 将包含祀DNA的样品引入反应位点。 The sample containing DNA introduced into the reaction worship sites. 如果祀DNA中存在与探针互补的等位基因,则进行扩增,由此产生在上述检测中所描述的可检测信号。 If there is complementary to the probe DNA of Si allele is amplified, thereby generating a detectable signal is detected as described above. 根据所获的差别信号,可W确定多态性位点处的核巧酸身份。 According to the obtained difference signal, W can determine the identity of nuclear Qiao acid polymorphism at the site. 如果两个信号都被检测到,则两个等位基因都存在。 If the two signals are detected, then both alleles are present. 如上述溫度控制部分所述,进行反应期间的热循环。 As part of the above-described temperature control, thermal cycling during the reaction.

[0079] 基因表达分析包括:测定在特定细胞中表达一种或多种基因的水平。 [0079] Gene expression analysis comprising: determining the expression levels of one or more genes in a particular cell. 运种测定可W是定性的,但通常是定量的。 W transported species determination may be qualitative, but usually quantitative. 在差别基因表达分析中,将一种细胞(例如受试细胞)中的基因水平与另一种细胞(对照细胞)中相同基因的表达水平进行比较。 In the differential gene expression analysis, the expression level of one cell (e.g., test cell) level of the gene and another cell (control cell) are compared in the same gene. 可W做出非常多的运种比较。 W can make a lot of kinds of comparison operation. 示例包括但不限于:健康细胞和病变细胞之间的比较;用一种药物治疗的个体的细胞和未治疗个体的细胞之间的比较;接触和未接触具体毒剂的细胞之间的比较,等等。 Examples include, but are not limited to: Comparison between healthy cells and diseased cells; the comparison between the individual cells of the individual cells with a drug treated and untreated; compare the contact and not in contact with specific agents between cells, etc. Wait. 在受试和对照细胞中表达水平不同的那些基因可W用作治疗的标记物和/或祀点。 Those different levels of gene expression in the test and control cells can be used as a therapeutic marker W and / or the point of worship. 例如, 如果在病变细胞而非健康细胞中发现某组基因被上调,则运些基因可W用作疾病的标记物并且有可能用作诊断测试的基础。 For example, if you find a group of genes are upregulated in diseased cells but not healthy cells, then transport these genes can be used as a marker of disease W and may be used as the basis of a diagnostic test. 运些基因也可W是祀点。 These genes may also be transported W is the point of worship. 治疗该疾病的方案可包括导致运些上调基因表达降低的步骤。 Treatment of the disease may include a program of steps leading gene expression decreased shipped some increase.

[0080] 本发明实施方式中所使用的微流体器件的设计有利于各种基因表达分析。 [0080] The microfluidic device design embodiment of the present invention is used in a variety of gene expression analysis in favor. 因为运些器件包含大量的反应位点,所W可同时测定大量的基因和/或样品。 These devices contain a large number of transport because of the reaction sites, the W can be determined simultaneously a large number of genes and / or samples. 采用盲端流体通道器件,可W同时测定成百上千种基因的表达水平。 Blind-side fluid channel device can W simultaneously determining the expression levels of hundreds of genes. 运些器件也有利于差别基因表达分析。 These devices also help transport differential gene expression analysis. 在使用矩阵设计的情况下,获自健康细胞的样品可W在一条流体通道中进行测试,而来自病变细胞的样品则在紧邻的通道中运行。 In the case of using the design matrix, a sample obtained from a healthy cell can be tested in a W in the fluid passage, and a sample of diseased cells from the run in the immediate vicinity of the passage. 运种特征使检测变得容易,并且提高了结果的精确度,因为两个样品同时在同一器件上运行并且处于相同的条件下。 Yun species feature makes it easy to detect, and improve the accuracy of the results, because the two samples were run simultaneously on the same device and in the same condition.

[0081] 根据本发明的各实施方式,也使用各种矩阵器件或W阵列为基础的器件。 [0081] According to the embodiments of the present invention may also use a variety of matrix devices or W-array-based devices. 某些运样的器件包括:(i)第一类多个流体通道,形成于弹性基板中;(ii)第二类多个流体通道, 形成于弹性基板中并与第一类多个流体通道相交从而限定了反应位点的阵列;(iii)多个隔离阀,设置在第一类和第二类多个流体通道内,可用于将各反应位点内的溶液与其它反应位点处的溶液隔离开;W及(iv)多个保护通道,它们覆盖在一条或多条流体通道和/或一个或多个反应位点上,W防止溶液蒸发。 Some kind of transport device comprising: (i) The first plurality of fluid passages formed in the elastic substrate; (ii) The second category plurality of fluid channels formed in the elastomeric substrate of the first type and a plurality of fluid channels intersect thereby defining an array of reactive sites; (iii) a plurality of isolation valve provided in the first class and the second class of a plurality of fluid channels for the solution of the reactive sites within the remainder of the reaction at the site isolated solution; W and (iv) a plurality of guard channels, which cover the one or more fluid channels and / or one or more reactive sites, W to prevent evaporation of the solution. 上述器件可用于许多不同类型的反应,包括那些设及溫度调节的反应(例如,核酸分析的热循环)。 The devices can be used in many different types of reactions, including those regulating the reaction temperature and the set (e.g., thermocycling of nucleic acid analyzes).

[0082] 一些微流体器件使用一种常被称为"盲端通道"或"盲端填充"的设计,运种设计具有多个盲端通道,盲端通道是具有一个死端或隔离端使得溶液只能从该盲端通道的一个端进出的运样一种流体通道(即盲端通道没有单独分离的入口和出口)。 [0082] Some microfluidic devices using a process commonly referred to as "dead-end channel" or "The Blind Side fill" design, transportation design having a plurality of types of blind-ended channels, blind-ended channel having a dead-end or end such isolation solution from only one end of the blind end of the channel and out of a fluid sample transport channel (ie the blind end of the channel is no separate separate entrance and exit). 运些器件只需要给每一个盲端通道配单个阀,W便隔离盲端通道的区域从而形成隔离的反应位点。 These devices run just to give each a dead-end channel with a single valve, W will isolate the blind end of the channel region so as to form an isolated reaction site. 在运种类型的器件的制造过程中,用于进行分析的一种或多种试剂被沉积在反应位点处,由此使输入和输出的次数显著减小。 The number of one or more agents in the manufacturing process operation types of devices, for analysis in the reaction is deposited at the site, whereby the input and output significantly reduced. 另外,运些盲端通道连接到互连的通道网络,使得可W从单个或有限个样品输入填充所有的反应位点。 In addition, the transport of these dead-end channels are connected to interconnected network of channels, making it possible to fill all W reactive sites from a single or a limited number of sample input. 因为输入和输出复杂性减小了并且只使用单个阀来隔离每一个反应位点,所W可用于反应位点的空间增大了。 Because the input and output complexity is reduced and the use of only a single valve to isolate each reaction site, the W space available for reaction sites is increased. 由此,运些器件的特征意味着, 每一个器件可W包括大量的反应位点(例如,多达上万个),并且可W实现高反应位点密度(例如,每平方厘米超过1,000-4, 000个反应位点)。 Thereby, the transport characteristics of these devices means that each device can include a large number of W reactive sites (e.g., up to tens of thousands), and at the W high reaction site densities (e.g., more than one per square centimeter, 000-4, 000 reaction sites). 和常规的微流体器件相比,上述运些特征也直接地使器件的尺寸显著减小。 And compared to conventional microfluidic device, said transport some features may also direct that the size of the device is significantly reduced.

[0083] 本文所掲示的其它微流体器件使用矩阵设计。 [0083] As used herein, other microfluidic devices kei shown using a matrix design. 通常,运种类型的微流体器件使用多个相交的水平和垂直流体通道,W便在相交的点处定义反应位点的阵列。 Typically, the types of transport microfluidic device using a plurality of intersecting horizontal and vertical fluid passage, W will define the array at the point of intersection of the reactive sites. 由此,运种设计的器件也具有阵列或反应位点;然而,在使用的运种设计时,仍然有大量的样品输入和相应的样品输出来供给大量的样品。 Thus, the devices shipped kinds of design also has an array or reaction site; however, in the use of transport kinds of design, there are still a large number of sample input and output samples corresponding to supply large numbers of samples. 被称为可切换式流体阵列构架的阀系统能够使溶液选择性地流过水平或流体通道,由此允许对矩阵中的各种流体通道进行可切换的隔离。 Is called switchable fluid valve array system architecture enables the solution to selectively flow through horizontal or fluid passage, thereby allowing for a variety of matrix isolation fluid channel switchable. 因此,尽管盲端通道被设计成在不同的条件下用有限数目的样品来进行大量的分析,但是矩阵器件被构建成在有限的条件下分析大量的样品。 Thus, while the blind end of the channel is designed with a limited number of samples for analysis in a lot of different conditions, but the matrix device is constructed such that, under conditions of limited analysis of large numbers of samples. 其它器件都是运两个通用设计类型的混合。 Other devices are shipped mixing two types of universal design.

[0084] 其它微流体器件是整体分隔器件值ID),在下列文献中对此进行了描述:PCT公报WO 2004/089810;美国专利申请10/819088,作为US 20050019792 予W公布;2006 年6 月2日提交的共同待批共同转让的专利申请No._且题为"Analysis using microfluidic partitioning devices"(代理人档案标号020174-012400),运些文献全部引用在此作为参考。 [0084] Other microfluidic device is an integral component values separated ID), in the following documents which are described: PCT Publication WO 2004/089810; U.S. Patent Application 10/819088, published as US 20050019792 to W; June 2006 2 filed co-pending commonly assigned patent application No._ and entitled "Analysis using microfluidic partitioning devices" (Attorney Docket numeral 020174-012400), shipped some literature reference in its entirety herein by reference. 通过使用整体分隔器件,样品可W被分隔到多个隔离的反应室中,并且在每一个室内同时执行多个反应。 By using the integral separator device, the sample W can be separated into a plurality of isolated reaction chamber and executed simultaneously in each of a plurality of the reaction chamber.

[00化]本文所描述的微流体器件进一步还具有下列部分特征:使用由弹性材料制成的各种组件,比如流体通道、控制通道、阀和/或累。 [Of 00] The microfluidic device described herein has the following further characteristic part: various components of an elastic material, such as fluid channels, control channels, valves and / or tired. 在一些情况下, In some cases,

[0086] 基本上整个器件都是由弹性材料制成的。 [0086] substantially the entire device is made of an elastic material. 结果,和用塑料或W娃为主的材料构成的常规微流体器件相比,上述器件在形状和功能方面具有显著的不同。 As a result, conventional microfluidic device using a plastic material or a W-based baby compared with the above-described device significantly different in terms of shape and function. 根据本发明的各实施例,所提供的反应室的数目可W有所不同。 According to various embodiments of the present invention, the number provided by the reaction chamber W may vary.

[0087] 运些器件的设计能够使它们与许多不同的加热系统相结合地使用。 [0087] These devices are designed to be able to transport them with many different heating systems used in combination. 由此,运些器件可用于进行需要溫度控制的各种不同的分析。 Thus, these devices can be used for various transportation need for temperature control of the different analyzes. 另外,适合于用在加热应用中的微流体器件可W包括另一种设计特征,W便使反应位点处样品的蒸发最小化。 Also, suitable for use in heating applications in microfluidic device may include another design feature W, W will make the evaporation of the reaction at the site of the sample is minimized. 运种类型的器件通常包括许多形成于弹性器件内的保护通道和/或储液囊或室,通过它们可W使水流入从而增大构成该器件的弹性材料内的水蒸气气压,由此减小样品材料在反应位点处的蒸发。 Type transport device comprises a plurality of generally resilient means formed within the protection channel and / or reservoir or capsule chamber, through which at the W thereby increasing the water into water vapor pressure of the elastomeric material within the device, thereby reducing small sample material is evaporated in the reaction at the site.

[0088] 在另一个实施方式中,可用溫度循环器件来控制微流体器件的溫度。 [0088] In another embodiment, a temperature cycling device can be used to control the temperature of the microfluidic device. 较佳地,微流体器件将用于与微流体器件热接触。 Preferably, the microfluidic devices will be used in contact with the heat microfluidic device. 在基板材料支撑微流体器件的情况下,如载玻片或载板(如塑料托架)底部,可W在载板或载玻片的区域中形成窗口,使得微流体器件(最好是具有弹性模块的器件)可W直接接触溫度循环器件的加热/冷却模块。 In the microfluidic device substrate material supporting the case, such as a glass slide or plate carrier (e.g., plastic carriers) at the bottom, at the W window formed in the region of the carrier plate or glass slide such that the microfluidic device (preferably having a elasticity module device) W can be in direct contact with the temperature cycle of the device heating / cooling module. 在较佳的实施方式中,加热/冷却模块具有形成于其中的槽,运些槽和真空源相连,用于对该微流体器件施加吸入力,最好对发生反应的位点邻接的部分施加吸入力。 In a preferred embodiment, the heating / cooling module has a groove formed therein, the groove and some transport vacuum source for applying a suction force to the microfluidic device, preferably on the part of the reaction sites adjacent applying suction force. 或者,可W将刚性导热盘接合到微流体器件,然后与加热和冷却模块配对W便有效地进行热传导。 Alternatively, W rigid thermal plate is bonded to the microfluidic device, and then heating and cooling modules will be paired W efficient heat conduction.

[0089] 某些器件的阵列格式意味着运些器件可W实现高通量。 [0089] array format means that some devices these devices can be transported W high throughput. 总的说来,高通量和溫度控制能力使运些器件可用于进行大量核酸扩增(例如,聚合酶链反应(PCR))。 In general, high throughput and temperature control capabilities so that these devices can be used to transport a large number of nucleic acid amplification (eg, polymerase chain reaction (PCR)). 本文将详细讨论运种反应,W说明该器件的实用性,尤其是它们在需要溫度控制的任何反应中的应用。 This article will discuss the types of transport reactions, W illustrate the usefulness of the device, in particular, it is that they require any reaction temperature controlled applications. 然而,应该理解,运些器件并不限于运些特定应用。 However, it should be understood that these devices run some application-specific operation is not limited. 运些器件可用于各种其它类型的分析或反应之中。 These devices can be used to transport a variety of other types of analyzes or reactions being. 运些示例包括:分析蛋白质-配体相互作用;W及分析细胞与各种化合物之间的相互作用。 Transported some examples include: analysis of protein - ligand interactions; and analysis of the interaction between the W cells with various compounds. 在本说明书中提供了更多的示例。 Additional examples are provided in this specification.

[0090] 本文所掲示的微流体器件通常至少部分地由弹性材料构成,并且由单层和多层软光刻(MSL)技术和/或牺牲层包裹方法(例如,参照化ger等人,(2000) Science 288 : 113-116;化及PCT公报WO 01/01025,它们全部引用在此作为参考)。 [0090] As used herein, kei microfluidic device shown generally at least partially made of an elastic material, and the monolayer and multilayer soft lithography (MSL) techniques and / or sacrificial layer coating process (e.g., refer to provisions of ger et al., ( 2000) Science 288: 113-116; and PCT Publication of WO 01/01025, all of which are hereby incorporated by reference). 通过使用运些方法, 微流体器件可W被设计成,溶液流过该器件的流体通道的过程至少部分程度受到控制,其中用弹性膜或区段使一个或多个控制通道和流体通道分隔开。 By using these transportation methods, microfluidic devices can be designed to be W, the solution flowing through the fluid passage of the device during the at least partially under control, wherein an elastic membrane or a section of one or more control channels so that the fluid passage and the partition open. 通过向控制通道施加作用力,该膜或区段可W弯曲到和控制通道相关联的流体通道中或从该流体通道中缩回来。 By applying a force to the control channel, or a segment of the film can be bent W fluid channel to control channel associated or withdrew from the fluid passage. 通过控制上述膜弯曲到流体通道中的程度或者控制上述膜从该流体通道中缩回来的程度,在流体通道中的溶液流动可W放缓或完全被阻挡。 Bending by controlling the membrane into the fluid passage for controlling the degree or extent to which the film is retracted from the fluid passage, the solution flow in the fluid passage at the W slowed or completely blocked. 通过使用运种类型的控制通道和流体通道的组合,便可W制备各种不同类型的阀和累W便调节溶液流动,在下列文献中对此有更多的细节描述:Unger 等人,(2000) Science 288 :113-116 ; W及PCT 公报W0/02/43615 和WO 01/01025。 Using a combination of types of transport channels and control fluid channels, W can be prepared in various types of valves and tired W will adjust the solution flow in the following documents which have a more detailed description: Unger et al., ( 2000) Science 288: 113-116; W and PCT Publication W0 / 02/43615 and WO 01/01025.

[0091] 本文所提供的器件包括运种累和/或阀,W便选择性地隔离允许试剂发生反应的那个反应位点。 [0091] The device provided herein include species transport tired and / or valves, W will allow selective isolation reagent that reacts reaction site. 或者,使用不带累和/或阀的器件,运些器件使用压力驱动流或聚合过程来关闭合适的通道,并由此选择性地隔离反应位点。 Or, do not implicate the use and / or valve devices, these devices shipped using pressure-driven flow or polymerization process to close the appropriate channel, and thereby selectively isolate the reactive sites. 反应位点可W被定位到该器件内大量不同位置中的任何位置。 W reactive sites can be targeted to a number of different positions within the device in any location. 例如,在一些矩阵型器件中,反应位点被定位到一组流体通道的相交处。 For example, in some matrix-type devices, the reaction site is located at the intersection of a plurality of fluid channels. 在盲端通道器件中,反应位点被定位到盲端通道的一端。 In the blind-side channel devices, the reaction site is located to one side of the blind end of the channel.

[0092] 如果该器件用于溫度控制反应(例如,热循环反应),则如下文更详细的描述,弹性器件通常被固定到托架(例如载玻片)上。 A more detailed description of [0092] If the device is used for controlling the reaction temperature (e.g., thermocycling reactions), then below, the elastic devices are generally carrier (e.g., a glass slide) is fixed to the upper. 所得的结构可W接着被放到溫度控制板上, W便控制各个反应位点处的溫度。 The resulting structure may then be placed in W temperature control board, W will control the temperature of each reactor at the site. 在热循环反应的情况下,该器件可W被放到许多热循环板上。 In the case of thermal cycling reactions, the device can be placed in a number of thermal cycles W plate.

[0093] 因为运些器件均由相对而言具有光学透明性的弹性材料制成,所W在微流体器件上基本上任何位置处,通过使用各种不同的检测系统,都可W很容易地监控各种反应。 [0093] Because the operation of these devices by a relatively resilient material having optical transparency, the W at substantially any location on the microfluidic device, by using a variety of detection systems, can readily W monitoring a variety of responses. 然而,通常,都是在反应位点自身处进行检测(例如,在包括流体通道交点的区域或在流体通道的盲端)。 However, in general, are detected (for example, including a fluid passage in the intersection area or blind-side fluid passage) in the reaction at the site itself. 上述器件由基本上透明的材料制成运一事实也意味着,某些检测系统可W和目前尚不能与常规娃基微流体器件一起使用的那些器件一起使用。 By the fact that said device shipped a substantially transparent material also means that certain detection systems at the W and those devices not currently conventional baby-yl use with microfluidic device used in conjunction. 通过使用被嵌入上述器件中的检测器,或者通过使用与上述器件分离但与待测器件区域对准的检测器,便可W实现检测。 By using the above device is embedded in the detector, or a separate but aligned with the region of the device under test and the detector through the use of the above device, the detection can be realized W.

[0094] 使用矩阵设计的器件通常具有多个垂直的和水平的流体通道,运些流体通道彼此相交W便形成多个结的阵列。 [0094] using a matrix design devices typically have a plurality of vertical and horizontal fluid passage, some transport fluid passage intersecting each W is formed an array of multiple junctions. 因为不同的样品和试剂(或试剂组)可W被引入每一个流体通道,所W可W按高通量形式在相对大量的反应条件下测试大量的样品。 Because of different sample and reagent (or set of reagents) is introduced at the W each fluid passage, which at the W by W at a high throughput format relatively large number of reaction conditions, a large number of test samples. 由此,如果不同的样品被引入M个不同的垂直流体通道中的每一个通道之内并且不同的试剂(或试剂组)被引入N个水平的流体通道的每一个通道之内,则可W同时进行MxN个不同的反应。 Thus, if a different sample is introduced and the different M different vertical flow channels of a channel of each of the reagent (or set of reagents) is introduced into each channel of the N channels of the fluid level can be W MxN simultaneously different reactions. 通常,矩阵器件包括阀,允许对垂直和水平的流体通道进行可切换的隔离。 Typically, matrix devices include valves that allow for horizontal and vertical fluid channel switchable isolation. 运些阀定位成允许选择性地只流过垂直的流体通道,或只流过水平的流体通道。 These valves positioned to permit operation to selectively flow only through the vertical fluid passage, or horizontal flow only through the fluid passage. 因为该类型的器件具有和样品类型和数目选择、试剂数目和类型选择等有关的灵活性,所W运些器件可很好地适用于进行许多特定的分析,在运些特定的分析中,人们想要根据相对大量的反应条件来筛选大量的样品。 Because this type of device has about flexibility and sample type and number of options, the number and type of agent selection, the W transported these devices is well adapted to carry out a number of specific analysis, in particular analysis of some transportation, people according to want a relatively large number of reaction conditions for screening large numbers of samples. 矩阵器件可W选择性地包括保护通道,W便防止样品和试剂的蒸发。 W matrix devices can optionally include the protection channel, W will prevent the evaporation of sample and reagent.

[0095] 一些高密度矩阵设计利用了微流体器件各层之间的流体连通孔,W便使整个器件中的控制线和流体线交织起来。 [0095] Some high-density matrix design utilizes fluid communication hole microfluidic device between the layers, W will make the entire device control lines and fluid lines intertwined together. 例如,通过使双层弹性模块的每一层中都具有流体线,密度更高的反应室排布是可行的。 For example, by double elastic module of each layer, having a fluid line, the higher the density of the reaction chamber arrangement is feasible. 对于本领域的技术人员而言,很明显,多层器件允许流体线彼此在上或在下相交,同时不处于流体传递的状态中。 Skilled in the art it will be obvious, multilayer device allows fluid lines intersect each other or lower, while not in a state of fluid transfer. 例如,在特定的设计中,第一层中的试剂流体通道通过一个孔连接到第二层中的试剂流体通道,而第二层中还具有样品通道,样品通道和试剂通道分别终结于样品和试剂室。 For example, in a particular design, the first reagent layer is connected through a fluid passage hole to the second fluid passage reagent layer, the second layer further has a sample channel, channel samples and reagents at the sample channel end, respectively, and reagent chamber. 样品室和试剂室通过一个连接通道而进行流体传递,该连接通道具有相关的连接阀,W便控制反应室的每一个室之间的流体传递。 Sample chamber and the reagent chamber through a connecting passage and fluid transfer, the connection channel with an associated connecting valve, W will control fluid to pass each chamber between the reaction chamber. 在使用过程中,该连接首先闭合,然后从试剂入口将试剂引入试剂通道中,并且从样品入口将样品引入样品通道中。 During use, the connection is closed first, and then introduced into the reagent from the reagent inlet channel reagent, and the sample from the sample inlet for sample introduction channel. 然后,密封阀闭合,W便使反应室和其它室隔离开。 Then, the sealing valve is closed, W will be the reaction chamber and the other chamber isolated. 一旦反应室被隔离, 则打开连接阀W便使样品室和试剂室进行流体传递,从而可W发生想要的反应。 Once the reaction chamber is isolated, it will open a connection to the sample chamber valve W and the reagent chamber in fluid transfer, which can occur W desired reaction. 本领域的技术人员将认识到许多变体、修改和替换。 Those skilled in the art will recognize that many variations, modifications and substitutions.

[0096] 相应地,用于微流体器件的特定设计提供了一种微流体器件,它适用于使M种不同的样品和N种不同的试剂进行反应,该器件包括:多个反应室,每一个反应室包括样品室和试剂室,样品室和试剂室通过连接通道进行流体传递,该连接通道具有相关的连接阀,用于控制样品室和试剂室之间的流体传递;多个样品入口,都和样品室相通;多个试剂入口, 都和试剂室相通;其中样品入口或试剂入口之一通过小孔分别与样品室之一或试剂室之一进行流体传递。 [0096] Accordingly, the particular design of the microfluidic device to provide a microfluidic device, so that it applies to different kinds of samples M and N different reagent reaction, the device comprising: a plurality of reaction chambers, each a reaction chamber including a sample chamber and a reagent chamber, the sample chamber and a reagent chamber in fluid passing through the connecting passage, the connecting passage is connected with an associated valve, for controlling fluid flow between the sample chamber and a reagent chamber transfer; a plurality of sample inlet, and the sample chamber are connected; a plurality of reagent inlet, and the reagent chamber are interlinked; wherein the sample inlet or one reagent inlet orifice through each one of the sample chamber of the reagent chamber or fluid delivery. 某些实施例包括:在多层结合所构成的弹性模块中形成多个反应室,并且连接阀是可弯曲的膜;使样品入口通过样品通道和样品室相通,使试剂入口通过试剂通道和试剂室相通,样品通道的一部分和试剂通道的一部分定向成大约彼此平行,并且各自具有相关联的密封阀,用于控制所通过的流体传递过程;使与样品通道相关联的阀和与试剂通道相关联的阀通过共用的密封控制通道彼此相通;使密封共用控制通道沿着大约和样品通道或试剂通道正交的线定位。 Some embodiments include: elasticity module formed in a multilayer combination composed plurality of reaction chambers, and the valve is connected to the flexible membrane; sample channel through the sample inlet and a sample chamber communicating, through the reagent inlet passage and Reagents Reagents room interlinked, part of the channel portion of the sample and reagent channel is oriented approximately parallel to each other, and each having an associated sealing valve for fluid transfer through the process control; valve so that the channel associated with the sample and reagent channel associated with linked by a common sealing valve control channel communicated with each other; the seal along the common control channel and the sample channel or approximately orthogonal to the reagent channel line positioning.

[0097] 本发明各实施例中所使用的微流体器件可W进一步被集成到一种托架器件中, 运在共同待批、共同拥有的由化ger filed于2004年3月29日提交的美国专利申请60/557,715中进行了描述,该专利申请引用在此作为参考。 [0097] Examples of microfluidic devices used in embodiments of the present invention further W can be integrated into one kind of device bay, transported in co-pending, co-owned by the filed of ger on March 29, 2004 filed U.S. Patent application 60 / 557,715 are described in the patent applications are incorporated herein by reference. 化ger的托架提供了工作中的连续流体压力,W便保持阀闭合状态远离流体压力源(例如,房间空气压力)。 Ger bracket of the work provided continuous fluid pressure, W will be kept away from the valve closed the fluid pressure source (for example, room air pressure). 如上述专利申请中所述的,linger进一步提供了一种自动系统,用于充注并启动本发明的阀。 As described in the above patent application, linger further provides an automated system for filling and start the valve of the present invention. 在另一个较佳实施例中,用于充注蓄能器并启动阀的自动系统使用了一种具有滚筒的器件,该滚筒顶住微流体器件的一个或多个表面,其中该滚筒具有至少两个或多个与受控真空或压力源相通的端口,并且可W包括用于操纵微流体器件多个部分的机械部分,例如但不限于单向止回阀。 In another preferred embodiment, for filling the accumulator and start the automatic valve system uses a cylinder device having one or more of the drum against the surface of the microfluidic device, wherein the drum has at least two or more controlled vacuum or pressure source communicating port, and at the W include a plurality of portions for the mechanical part of the microfluidic device to manipulate, such as but not limited to a one-way valve.

[0098] 本发明各实施例中所使用的另一种器件提供了一种被用作基板的托架,用于使弹性模块稳定。 [0098] Another embodiment of the device used to provide a tray to be used as a substrate for the elastic module stability in all embodiments of the present invention. 较佳地,该托架具有一个或多个下面的特征:一个井或储液囊,它通过形成于托架中或与托架一起形成的至少一个通道而与弹性模块相通;蓄能器,它通过形成于托架中或与托架一起形成的至少一个通道而与弹性模块相通;W及流体端口,与弹性模块相通, 其中该流体端口最好可W接入自动真空或压力源(比如上述自动系统),其中自动源还包括一种具有端口的滚筒,它与上述流体端口配对,W便在用于将流体压力或真空施加到弹性模块的自动系统之间形成隔离的流体连接。 Preferably, the carrier has one or more of the following features: a fluid bag or reservoir well, it is formed by at least one channel in the bracket or the bracket is formed together with the elasticity module in communication; accumulator, it is formed by at least one channel together with the bracket or the bracket is formed in communication with the elasticity module; W and the fluid port, in communication with the elasticity module, wherein the fluid access port is preferably at the W automatic vacuum or pressure source (such as said automatic system), which also include automatic source port having a drum, with said fluid port pairing, W will be in for a fluid pressure or vacuum is applied to the isolation of the fluid connection is formed between the elastic automatic system module. 在某些实施例所用的器件中,自动源也可W 与一个或多个与托架相关联的蓄能器相通,W便充注并释放蓄能器中所维持的压力。 In certain embodiments of the device used, the automatic source can also be a W or accumulator with a plurality of brackets associated communication, W will charge and release the accumulator pressure maintained. 在某些实施例中,该托架可W进一步包括定位于该托架范围中接触微流体器件的一个区域,其中该区域由与该托架另一个部分不相同的材料制成,该区域材料的选择是为了改善热传导和热分布特性,运些与该托架的其它部分不相同。 In certain embodiments, the carrier may further comprise W positioned on the carrier in a range of the contact area of the microfluidic device, wherein the region made of another portion of the carrier material is not the same, the material in the region the choice is to improve the thermal conductivity and heat distribution, transport and some other parts of the bay are not the same. 用于改善热传导和热分布的较佳材料包括但不限于娃,最好是高度抛光的娃,比如半导体领域中可获得的抛光的晶片或从晶片中切割的一部分即忍片。 Preferred material for improving the heat conduction and heat distribution including but not limited to baby, the baby is preferably highly polished, polishing of semiconductor wafers, such as available in the art that is a part or forbearance piece cut from the wafer.

[0099] 如下文更全面地描述,本发明的各实施例使用一种热源,但不限于PCR热循环装置,它可能已从其原始制造状态中获得修改。 [0099] As described more fully below, embodiments of the present invention uses a heat source, but not limited to PCR thermocycler device, it may have been obtained in its original manufactured condition changes. 通常,该热源具有热调节部分,该部分与托架的一部分(最好是托架的热传导和热分布部分)配对,W便通过该托架的热传导和热分布部分向弹性模块提供热控制。 Typically, the heat source having a thermal conditioning section, a portion of the portion of the bracket (the bracket is the best heat conduction and heat distribution section) pairing, W will provide heat to the elasticity module through heat conduction and heat distribution control portion of the bracket. 在较佳的实施例中,通过将真空源施加到热源的热调节部分内所形成的一个或多个通道,便改善了热接触,其中形成多个通道W便接触该托架的热传导和热分布部分的表面,从而施加吸力并维持该托架的热传导和热分布部分的位置。 In the preferred embodiment, the vacuum source is applied by the heat source to the adjustment of one or more channels formed in the part, it improves the thermal contact, wherein a plurality of channels W will be contacted and the carrier thermal conduction heat distribution of the surface portion, so as to apply suction position and maintain thermal conduction and distribution portion of the heat carrier. 在较佳的实施例中,通过将热传导和热分布部分只附着于弹性模块上并且在热传导和热分布部分的边缘周围留下空隙W便减小由该托架造成的寄生热效应,该托架的热传导和热分布部分并不与该托架的其余部分进行物理接触,而是与该托架的其余部分和弹性模块相关联。 In the preferred embodiment, by the heat conduction and heat distribution section is only attached to the elastic module and heat transfer around the edge portions of the heat distribution and W leaving voids will reduce parasitic thermal effects caused by the the carrier, the carrier heat transfer and heat distribution part is not in physical contact with the rest of the bay, but with the rest of the carrier and the elastic module is associated. 应该理解,在本文所描述的本发明的许多方面,较佳的弹性模块可W用本文没有描述但本领域已知的任何微流体器件来替代,例如由美国加州Santa Clara的Affymetrix(R)公司或美国加州Mountain View的化Iiper公司所生产的器件GeneQiip(R)。 It should be understood that in many aspects of the present invention described herein, the preferred elasticity module W is not described herein but known in the art to replace any microfluidic device, for example, a Santa Clara, California in the Affymetrix (R) company or Mountain View, California-based company produces Iiper devices GeneQiip (R). 授权给Soane、 Pa;rce、Fodo;r、Wilding、Ekst;rom、如ake或Unger的美国专利描述了可W被替代的微流体器件或中尺度流体器件,因为本发明的弹性模块利用了上述溫度方面的优点和改进,例如吸入定位,减少到流体器件的其它区域的寄生热转移,运些在上文使用弹性模块的情况下进行过描述。 Licensed to Soane, Pa; rce, Fodo; r, Wilding, Ekst; rom, as described in U.S. Patent ake or Unger described can W be replaced microfluidic device or mesoscale fluidic devices, because the elastic module of the present invention uses the above the advantages and improvements terms of temperature, e.g., suction positioning, reducing to other areas of the fluid transfer device of parasitic heat, the case of transport modules above some elastic been described. 阳100] 通过使用本发明各实施例所提供的系统和方法,用384孔系统提高了通量。 Yang 100] By using the present invention, various embodiments of a system and method examples provided, with the 384 system improves throughput. 例如, 在一些实施方式中使通量W因数4、6、12、24或更大增加。 For example, in some embodiments, the manipulation factor 4,6,12,24 flux W or greater increase. 在产生通量增加的同时,还减小了各种操作的逻辑冲突。 Generating an increase in flux, it is also logical to reduce the conflicts of various operations. 此外,本发明各实施方式的系统和方法能够进行多个样品的多次试验。 In addition, the system and method of embodiments of the present invention can be performed repeatedly a plurality of test samples. 例如,在特定的实施方式中,采用96个样品和96次试验,提供了总共9, 216个数据点。 For example, in certain embodiments, the use of 96 samples and 96 tests, provided a total of 9, 216 data points. 在特定的示例中,运96次试验是化qMan 5'核酸酶试验的一部分。 In a particular example, the transportation is part of the 96 trials qMan 5 'nuclease tests. 阳101 ] 此外,本发明的各实施方式减小了反应体积。 Male 101] In addition, embodiments of the present invention reduces the reaction volume. 在本发明的各实施例中,所用反应体积为10皮升(10 12升)-100纳升(10 9升)。 In various embodiments of the present invention, the use of the reaction volume is 10 picoliters (1012 l) -100 nanoliter (109 liters). 在一些实施方式中,使用大于100纳升的反应体积。 In some embodiments, larger than 100 nanoliter reaction volumes. 仅作为示例,在一个实施方式中,本发明的方法和系统所用反应体积为10皮升、50 皮升、100皮升、250皮升、500皮升和I纳升。 Only as an example, in one embodiment, the method and system of the present invention is used in the reaction volume of 10 pl, 50 pl, pl 100, 250 picoliters and 500 picoliters I nanoliter. 在另选的实施方式中,使用2纳升、5纳升、10 纳升、20纳升、30纳升、40纳升、50纳升、75纳升和100纳升的反应体积。 In an alternative embodiment, the use of 2 nanoliters, 5 nanoliters, 10 nanoliter, 20 nanoliter, 30 nanoliter, 40 nanoliter, 50 nanoliter, 75 nanoliter and 100 nanoliter reaction volumes.

[0102] 根据特定微流体器件的几何形状和该微流体器件的大小W及流体连接通道和处理位点的排列情况,本发明的各实施例提供了范围不同的处理位点(或反应室)密度。 [0102] connection arrangement of channels and processing sites depending on the size W and fluid geometry specific microfluidic device and the microfluidic device, embodiments of the present invention provide a range of different treatment sites (or reaction chamber) density. 在一些实施例中,在使用本发明的方法和系统时,反应室密度大约从每平方厘米100个反应室到每平方厘米1百万个反应室。 In some embodiments, when the method and system of the present invention, the reaction chamber from a density of about 100 per square centimeter of the reaction chamber to the reaction chamber 1 million per square centimeter. 仅作为示例,根据本发明的各实施例,使用了每平方厘米有250个、1,000个、2, 500个、10, 000个、25, 000个、100, 000个和250, 000个反应室的反应室密度。 Only as an example, according to various embodiments of the present invention, the use of every square centimeter 250, 1,000, 2, 500, 10, 000, 25, 000, 100, 000 and 250, 000 the density of the reaction chamber of the reaction chamber. 在一些实施例中,使用了每平方厘米超过1,000, 000个反应室的反应室密度,尽管运不是本发明所要求的。 In some embodiments, the use of more than 1,000 per square centimeter, the density of the reaction chamber of the reaction chamber 000, although not the operation of the present invention as claimed. 阳103] 操作具有运种小反应体积的微流体器件可W减小试剂用量和样品用量。 Yang microfluidic device 103] operations have shipped species of small reaction volume W can reduce the amount of reagent and sample usage. 此外,本发明的一些实施方式提供了与实时定量PCR联用时可进行实时检测的方法和系统。 In addition, some embodiments of the present invention provides a method and system for real-time quantitative PCR and real-time detection can be associated with time. 使用运些系统和方法,可W为某些应用提供六个数量级的线性动态范围,并且使定量分辨率足够高W致于可W检测10纳升体积中亚纳摩尔级的巧光团浓度。 Transport systems and the use of these methods W may provide six orders of magnitude linear dynamic range for certain applications, and the quantitative resolution high enough to cause W to 10 W detect nanoliter volumes in sub-nanomolar concentration levels clever light group. 本领域的技术人员将会认识到许多改变、修改和替换。 Those skilled in the art will recognize that many changes, modifications and substitutions.

[0104] 用某些盲端通道类型的器件实现的方法包括:提供一种微流体器件,该微流体器件包括形成于弹性材料内的流体通道;W及多个盲端流体通道,它们与流体通道流体相连, 各盲端流体通道的末端区域限定了一个反应位点。 [0104] Certain Methods of the blind end of the channel type of device implemented method comprising: providing a microfluidic device, the microfluidic device comprising a fluid passage formed in the inside of the elastomeric material; the W and a plurality of blind-side fluid passage, and the fluid which a fluid passage connected to the end region of each blind end defines a fluid passageway reaction site. 至少一种试剂被引入各反应位点中,然后,在一个或多个反应位点处检测反应。 At least one reagent is introduced into each of the reaction sites, and then the reaction is detected at one or more sites of reaction. 该方法可W任选地包括加热反应位点内的至少一种试剂。 The method may optionally include W heating the at least one reagent reactive sites. 因此,一种方法可W包括:引入用于核酸扩增反应的各种成分;然后使运些成分热循环W形成扩增产物。 Thus, at the W A method comprising: introducing the various components for nucleic acid amplification reaction; then transported these components W of thermal cycling amplification product. 如下文更全面的描述那样,根据本发明各实施方式,提供了一种光学成像系统,它适用于表征某些微流体器件中发生的反应。 As more fully described above, according to embodiments of the present invention, there is provided an optical imaging system, which is suitable for characterizing certain reactions in the microfluidic device.

[01化]如图IA所示,本发明的一些实施例所提供的光学成像系统包括禪合到热控制模块的巧光成像系统。 [Of 01] As shown in Figure IA, some of the optical imaging system provided by the embodiments of the present invention comprises a thermal control module Zen bonded to the clever optical imaging system. 一些系统适用于收集来自几何形状为NxM的微流体忍片的数据。 Some systems applicable to collect data from the geometry of NxM microfluidic endure tablets. 在一些实施例中,N等于M。 In some embodiments, N is equal to M. 例如,本发明的各实施例使用具有48x48个反应室的微流体器件、具有96x96个反应室的微流体器件、W及其它几何形状的微流体器件。 For example, various embodiments of the present invention uses a reaction chamber having 48x48 microfluidic device, having a reaction chamber 96x96 microfluidic device, W, and other microfluidic device geometries. 在特定的实施例中,在具有96x96反应室几何形状的微流体器件中,使用了96个样品和96个试剂。 In a particular embodiment, the microfluidic device having a reaction chamber geometry 96x96 using 96 samples and 96 reagent. 对于本领域的技术人员而言,很明显,本发明各实施例所提供的方法和系统能够使一个平台执行多种应用。 Skilled in the art, it is obvious, embodiments of the present invention to provide a method and system of the embodiment enables a platform to perform a variety of applications. 阳106] 图IA是示出了本发明一实施例的光学成像系统的简化示意图。 Yang 106] FIG. IA is a simplified schematic diagram illustrating an optical imaging system according to an embodiment of the present invention. 如图IA所示,根据本发明各实施例,提供了光源242。 As shown in Figure the IA, in accordance with various embodiments of the present invention, there is provided a light source 242. 如下文更全面地描述,在本发明的一些实施例中,来自光源242的光线被用来诱导样品中的巧光。 As described more fully below, in some embodiments of the present invention, the light from the light source 242 is used to induce sample clever light. 在其它实施例中,化学发光被用作一种指示。 In other embodiments, chemiluminescence is used as an indicator. 根据该实施例,将添加、除去、或使用各种系统成分,运对本领域的技术人员而言是很明显的。 According to this embodiment, will be added, removed, or use various system components, shipped to the skilled artisan is obvious. 在各种实施例中,使用发光二极管(LED)、激光器、弧光灯、白识灯等光源。 In various embodiments, the use of a light emitting diode (the LED), a laser, an arc lamp, a white light source such knowledge. 运些源可W是多色的或单色的。 These sources can be transported W is a multi-color or monochrome. 在特定的实施例中,该光源具有第一光谱带宽。 In a particular embodiment, the first light source having a spectral bandwidth. 在特定的实施例中,该光源是白光源,可产生光谱范围介于约400纳米到约700纳米的光学福射。 In a particular embodiment, the light source is a white light source, can produce spectral range between about 400 nm to about 700 nm optical emission of blessing. 仅作为示例,在本发明的一些实施例中,从加州Novato市Sutter Instruments公司那里可W买到的Lambda LS 300W氣弧光灯被用作光源。 Only as an example, in some embodiments of the present invention, from the city of Novato, California, where the company can Sutter Instruments Lambda LS 300W W buy gas arc lamp is used as the light source. 对于本领域的技术人员而言,很明显,在可选实施例中能够使用光谱带宽更大或更小的其它光源。 Skilled in the art, it is quite clear that in an alternative embodiment, the spectral bandwidth can be used more or less of other light sources. 阳107] 图IA示出了激发滤光轮244。 Yang 107] Figure IA shows excitation filter wheel 244. 在一些实施例中(例如,光源为多色的实施例),使用激发滤光轮244,对光源242发出的光线进行光谱滤光。 In some embodiments (for example, the light source is polychromatic embodiment), using an excitation filter wheel 244, the light emitted from the light source 242 spectral filtering. 当然,也可W使用多个滤光片。 Of course, the use of multiple filters may also be W. 作为一个示例,在一个实施例中,激发滤光轮提供了许多光谱滤光片,每一个光谱滤光片透射能够从样品中激发出特定巧光的预定波长范围。 As an example, in one embodiment, the excitation filter wheel provides a number of spectral filter, a spectral filter for each transmission inspire a certain predetermined wavelength range clever light from the sample. 如图IA所示,激发滤光轮244禪合到计算机270,从而提供滤光片的计算机控制。 FIG. IA, the excitation filter wheel 244 Zen bonded to the computer 270, thereby providing a filter computer control. 在特定的实施例中,激发滤光轮提供了许多光谱滤光片: In a particular embodiment, the excitation filter wheel provides a number of spectral filter:

[0108] 滤光片1 :其中屯、波长为485nm,光谱带宽为20nm ; [0108] Filter 1: wherein Tuen wavelength of 485nm, spectral bandwidth of 20nm;

[0109] 滤光片2 :其中屯、波长为530皿,光谱带宽为20皿;W及 [0109] Filter 2: Tuen wherein a wavelength of 530 dish, spectral bandwidth of 20 dish; W and

[0110] 滤光片3 :其中屯、波长为580皿,光谱带宽为20皿。 [0110] Filter 3: Tuen wherein a wavelength of 580 dish, spectral bandwidth of 20 dish. 阳111] 对于本领域的技术人员而言,很清楚,本发明的各实施例并不限于运些特定的光谱滤光片,而是将针对特定的样品使用适于巧光处理的光谱滤光片。 Yang 111] skilled in the art, it is quite clear that the various embodiments of the present invention is not limited to these specific spectral filter operation, but the clever use of light adapted spectral filter processing for a particular sample sheet. 此外,尽管上述讨论设及滤光轮的使用,但是运并不是本发明所要求的。 Further, although the above discussion and the use of the filter wheel is provided, but the operation of the present invention is not required. 在可选的实施例中,光谱滤光片W除轮子W外的更多几何形状来设置。 In an alternative embodiment, the spectral filter wheel W W addition of more geometric shapes to the outer set. 例如,根据本发明的各实施例,可W包括落入滤光片托架中的光谱滤光片、电光滤光片、由执行器将其置入光路中的滤光片等。 For example, according to various embodiments of the present invention, W to embrace filter bay spectral filter, an electro-optical filter, by the actuator to filter into the light path and the like. 此外,在其它实施例中,光源是可调谐的激光器,可W发射适于激发巧光的预定波长的福射。 In addition, in other embodiments, the light source is a tunable laser can emit a blessing W radio adapted excitation light of a predetermined wavelength clever. 本领域的技术人员将会认识到许多变体、修改和替换。 Those skilled in the art will recognize that many variations, modifications and substitutions.

[0112] 如图IA所示,根据本发明的各实施例,提供了激发快口246。 [0112] As shown in Figure IA, according to various embodiments of the present invention, there is provided a fast excitation port 246. 在一些实施例中,在计算机270的控制下,操作该激发快口246, W便阻挡/透射由光源242产生并由激发滤光轮244进行光谱滤光之后的光信号。 In some embodiments, the computer 270 under the control of the operation of the fast excitation port 246, W feces barrier / transmission optical signal 242 generated by the excitation filter wheel 244 after spectral filtering by the light source. 根据运种应用,在样品被插入系统中或从系统中取出样品时W及在校准操作期间,激发源被阻挡。 According to transport a variety of applications, the sample is inserted into the system, or W and during calibration operations, the excitation source is blocked when the sample was removed from the system. 在一些实施例中,激发快口并不用在使用激光光源的实施例中,使用激光光源的实施例提供可替换的手段来使光源焰灭。 In some embodiments, the excitation fast mouth is not used in a laser light source embodiment of the use of embodiments of the laser light source to provide alternative means to make the flame light off.

[0113] 当激发快口处于打开位置时,光激发信号穿过光纤束248并被引导照射到忍片托架中所设置的微流体器件205上。 [0113] When the excitation fast mouth is in the open position, the optical excitation signal through the optical fiber bundle 248 and is directed onto the irradiated plate carrier tolerance set in the microfluidic device 205. 本发明的其它实施例使用石英光导、液体光导、其它扰频系统等,来增大照明均一性。 Other embodiments of the present invention is the use of quartz guide, liquid light guide, other scrambling systems, to increase the illumination uniformity. 如图IA所示,引导激发光信号,使其通过光照明装置250的反射、折射或其组合,到达微流体器件205的表面上。 FIG IA, the guide excitation light signal, so that the light reflected by the illuminating device 250, or a combination of refraction, to the upper surface 205 of the microfluidic device. 如图IA所示,微流体器件的照明是通过光照明装置250进行的。 FIG IA, the illumination of the microfluidic device 250 through the illumination means carried. 在其它实施例中,通过环形光,或通过收集光学系统的一部分(微流体器件和检测器260之间的光路),照明可W从器件的一个或多个侧面倾斜地禪合到该微流体器件。 In other embodiments, the light through the ring, or by collecting part (microfluidic device and the detector optical path 260 between) the optical system, lighting can W from one or more sides of the device obliquely Zen bonded to the microfluidic devices.

[0114] 在一些实施例中,在样品的二维面积上,用激发源产生的光线对微流体器件进行照明。 [0114] In some embodiments, the two-dimensional area of the sample, the light generated by the excitation source microfluidic devices for lighting. 在运些实施例中,提供了较大的视场,运能够执行许多设及到给时间分辨化学过程和反应成像的巧光应用。 In some embodiments, transport, provide a larger field of view, and transportation can be provided to perform many time-resolved chemical processes for reactions and clever light imaging applications. 作为一个示例,蛋白质热量测定和核酸控制过程的巧光成像都是可从本发明各实施例中获益的时间分辨过程。 As one example, the protein calorimetry and clever optical imaging of nucleic acid control process are available from each embodiment of the present invention to benefit time to implement resolution process. 在运些过程中,期望同时激发许多反应室中所提供的多个巧光样品,并且还期望同时收集由运些反应室中正发生的各种反应所产生的巧光信号。 In the transportation process it desirable to simultaneously trigger multiple clever light many samples provided in the reaction chamber, and also desirable while collecting all kinds of clever optical signal transported by the reaction of some of the reaction chamber is occurring arising. 在其它过程(例如巧光寿命成像)中,在简短的激发脉冲之后,就是巧光信号的检测(和分析),因为巧光信号从其初始水平随时间流逝而衰减。 In other processes (such as Qiao light lifetime imaging), after a short excitation pulse is detected (and analysis) of the optical signal clever, clever because the optical signal from its original level over time and decay. 本领域的技术人员将会认识到许多变体、修改和替换。 Those skilled in the art will recognize that many variations, modifications and substitutions.

[0115] 作为一个示例,核酸扩增过程通常包括祀DNA、耐热DNA聚合酶、两种寡核巧酸引物、脱氧核巧酸=憐酸(dNTP)、反应缓冲液和儀盐。 [0115] As one example, the nucleic acid amplification process typically includes worship DNA, thermostable DNA polymerase, two oligonucleotide primers acid clever, clever deoxyribonucleic acid = pity acid (dNTP), reaction buffer and salt meter. 一旦混合后,就将该反应置于热循环器中,该热循环器是一种使该反应在一系列不同溫度下保持不同时间的仪器。 Once mixed, then the reaction is placed in a thermal cycler, which is a thermal cycler The reaction was held at different times in a series of different temperatures instruments. 运一系列溫度和时间调节被称为一个扩增循环。 Operation temperature range and is known as a time adjustment amplification cycle. 在该反应中,每一个循环从理论上讲能使祀序列(扩增子)的量倍增。 In this reaction, each cycle allows worship sequences (amplicons) from the amount theoretically doubled. 10次循环从理论上讲使该扩增子乘W约为1000的因数;在大约几个小时中,20次循环能使该扩增子乘W大于1百万的因数。 10 cycles so that theoretically the amplicon is about 1000 W multiplier factor; in about a few hours, 20 cycles enables the amplicon multiplication factor W is greater than 1 million. 在一些应用中,需要在几秒到几分钟的时间内获得很大面积(例如,若干个平方厘米的数量级)的巧光成像数据。 In some applications it is desirable to obtain a large area in a few seconds to a few minutes (for example, several square centimeters of magnitude) of clever light imaging data.

[0116] 在本发明的一些实施例中,本发明各实施例所提供的方法和系统有利于在预定的时间周期内执行的图像拍摄过程。 [0116] In some embodiments of the present invention, the various embodiments of the present invention is a method and system for image capturing process examples provided in favor performed within a predetermined time period. 仅作为一个示例,在本发明的一个实施例中,提供了一种给微流体器件成像的方法。 As just one example, in one embodiment of the present invention, there is provided a microfluidic device for imaging methods. 该方法包括:在不到一分钟的时间帖中,通过使用图像检测空间区域,拍摄与微流体器件的至少确定数目的反应室相关联的空间区域的图像,因此空间区域的图像拍摄基本上不受缝合和/或扫描过程的影响。 The method comprising: in less than a minute in the post, by using the image detection spatial region, the microfluidic device taken at least an image of the spatial region determined number of reaction chambers are associated, and therefore the image pickup region of space substantially without affected sewn and / or scanning process.

[0117] 本发明的各实施例提供了用于图像拍摄的各种时间帖,范围从1毫秒到1分钟。 [0117] Embodiments of the present invention is to provide a variety of time signatures for image capture, ranging from 1 millisecond to 1 minute. 在一些实施例中,用于图像拍摄的时间帖大于1分钟。 In some embodiments, the time for post shots than 1 minute. 根据与微流体器件的反应室中所执行的各种过程相关联的发光特性,用于图像拍摄的时间帖将发生变化。 The light emitting properties of the reaction chamber and the various processes in microfluidic devices associated performed for time signatures captured image will change. 例如,在一个实施例中,时间帖是10ms、50ms、100ms、250ms、500ms、750ms或1秒。 For example, in one embodiment, the post time is 10ms, 50ms, 100ms, 250ms, 500ms, 750ms or 1 second. 在其它实施例中,时间帖是2 秒、5秒、10秒、15秒、20秒、30秒、40秒、50秒、或1分钟。 In other embodiments, the post time is 2 seconds, 5 seconds, 10 seconds, 15 seconds, 20 seconds, 30 seconds, 40 seconds, 50 seconds or 1 minute. 当然,时间帖将取决于特定的应用。 Of course, the time signature will depend on the particular application.

[0118] 在一些实施例中,图像拍摄过程是按同步方式来执行的,从而可W同时拍摄确定数目的反应室的图像。 [0118] In some embodiments, the image capturing process is performed by a synchronized manner, thereby W concurrently with the images to determine the number of the reaction chamber. 作为一个示例,在示例性的PCR过程中,微流体器件维持92C 15秒。 As an example, in the example of the PCR process, the microfluidic device to maintain 92 C 15 sec. 接下来,该微流体器件维持60C 60秒。 Next, the microfluidic device to maintain 60 C 60 sec. 在一分钟的循环周期内重复加热和冷却,并且进行许多次循环。 In repeated heating and cooling cycles of one minute, and that many cycles. 通过使用本发明的各实施方式,同步地获得了微流体器件中存在的确定数目的反应室的图像,同时运些反应室根据位置保持在均匀的溫度下。 By using the embodiments of the present invention, the reaction chamber synchronously acquired image to determine the number present in the microfluidic device, while the reaction chamber in accordance with the position of some transport maintained at a uniform temperature. 例如,在微流体器件保持在溫度60C的情况下,使用30秒曝光,便可W获得整个微流体器件的二维图像。 For example, in the microfluidic device is kept at a temperature of 60 C case, a 30-second exposure, you can get the entire microfluidic device W two-dimensional image. 和光栅扫描或缝合系统相比,本领域的技术人员应该理解本发明所提供的诸多益处,因为在光栅扫描或缝合系统中微流体器件第一部分(例如,左上象限)中的反应室的图像是先于微流体器件第二部分(例如,右下象限)中的反应室的图像而获得的。 And rasterization or suturing system compared to those skilled in the art will appreciate the many benefits provided by the present invention, since the raster scanning system or suturing microfluidic device of the first portion (e.g., upper left quadrant) in the image is the reaction chamber microfluidic device prior to the second portion (e.g., lower right quadrant) of the image obtained by the reaction chamber.

[0119] 在其它实施方式中,在不到一分钟的时间帖内,获得了确定数目的反应室的多张图像。 [0119] In other embodiments, in less than a minute posts obtained to determine the number of reaction chambers of multiple images. 作为运些实施例的一个示例,在一个特定的实施例中获得了与多个巧光团相关联的多张图像。 As an example run some embodiments, in one particular embodiment, a plurality of received light clever group associated with multiple images. 在微流体器件保持在溫度60C下的60秒时间周期内,可W针对=种不同的巧光团(例如,R〇x™、Vic⑧和Fam™),获得使用20秒曝光的立张连续的图像。 Within the microfluidic device is maintained at a temperature of 60 C for 60 seconds under the time period, the W = for different kinds of clever light group (for example, R〇x ™, Vic⑧ and Fam ™), obtained using a 20-second exposure of Zhang Li continuous image. 当然,根据该应用,曝光时间可W更短,甚至短到1秒或更短。 Of course, according to the application, the exposure time may be shorter W, even as short as one second or less. 通过使用运些图像,可W计算并分析不同的巧光比。 By using these image transport, W can be calculated and analyzed different light than clever. 当然,根据巧光发射的强度,可W修改各种巧光团的曝光次数,W适应特定的应用。 Of course, depending on the intensity of light emission clever, W can modify various impressions clever light group, W suit a particular application. 运样,本发明的各实施例可W在微流体器件保持恒定溫度的同时在多个光谱波带中给微流体器件成像。 Sample transport, various embodiments of the present invention may be W microfluidic device while maintaining a constant temperature in a plurality of spectral band of imaging to the microfluidic device. 像上述示例所示,恒定的溫度可W是包括循环溫度处理的PCR过程的一部分。 As shown in the example above, a constant temperature W is part of the process include PCR cycle temperatures processed.

[0120] 本发明各实施例所提供的方法和系统也适合执行并分析化学发光过程。 Example methods and systems provided by various embodiments [0120] The present invention is also adapted to perform and analyze chemiluminescent process. 在运些过程中,各种反应发生在第一时间标度,而化学发光过程的图像则是在第二时间标度获得的。 In the transportation process, various reactions take place at the first time scale, and the image chemiluminescent process is the second time scale available. 在特定的过程中,第二时间标度小于第一时间标度。 In a particular process, the second time scale than the first time scale. 因此,当感兴趣的反应室中的样品已经反应了相等的时间量时,本发明的各实施例适于拍摄化学发光过程的同步图像。 Thus, when the reaction of interest in the sample chamber has a reaction equal amount of time, various embodiments of the present invention is suitable for shooting a chemiluminescent process synchronized image. 在运些过程中,提供了包括样品溫度循环的溫度控制,而在其它实施例中,反应室保持在恒定的溫度下。 In the course of these operation, it provides a temperature control comprises temperature cycling of the sample, while in other embodiments, the reaction chamber is maintained at a constant temperature. 阳121] 如图IA所示,根据本发明的各实施例,提供了热控制器240,也被称为溫度控制器240。 121] As shown in FIG Yang the IA, in accordance with various embodiments of the present invention, there is provided a thermal controller 240, the temperature controller 240 is also known. 改变精密化的许多不同的选择可用于控制微流体器件的选定区域内的溫度或整个微流体器件的溫度。 Change the precision of a number of different options can be used to control the temperature of the microfluidic device temperature within the selected area or the entire microfluidic device. 因此,如本文所使用的那样,术语溫度控制器广泛地指代一种器件或元件,它能够调节整个微流体器件的溫度或微流体器件的一部分内(例如,在特定的溫度区域内或微流体器件的通道矩阵的一个或多个结处)的溫度。 Thus, as used herein, the term broadly to refer to the temperature controller A device or element, it can be adjusted within a portion of the entire microfluidic device or the temperature of the microfluidic device (e.g., within a specific temperature range or slightly one or more fluid junction device channel matrix) temperature. 阳122] 图IC是示出了本发明一实施例的热控制器件的简化示意图。 Yang 122] Figure IC is a simplified schematic diagram illustrating a thermal control device according to an embodiment of the present invention. 如图IC所示,微流体器件205包括样品阵列206。 IC shown in FIG, microfluidic device 205 comprises an array of 206 samples. 对于本领域的技术人员而言,很明显,尽管图中样品阵列206 是一维的,但是本发明各实施例提供=维样品阵列。 For those skilled in the art, it is quite clear that, while the figure is one-dimensional array of samples 206, but embodiments of the present invention provides dimensional array of samples =. 作为一个示例,在本发明各实施例所使用的一些微流体器件中,反应室的阵列和流体连接通道延伸到该图所在的平面中。 As an example, the microfluidic device in some embodiments of the present invention is used, the reaction chamber arrays and fluidly connected channels extend to the plane of the diagram is located. 包括反应室在内的微流体器件的各种元件具有第=维度即深度。 A reaction chamber including a variety of elements, including a microfluidic device having a first dimension i.e., depth =. 微流体器件205由托架207支撑, 托架207由托架立柱208支撑。 Microfluidic device 205 supported by a bracket 207, the bracket 207 by a bracket 208 supporting columns. 微流体器件或忍片底层209禪合到托架207和集成热传播装置(MS) 241。 Microfluidic device or forbearance sheet bonded to the bottom of the bracket 207 209 Zen and integration of heat propagation means (MS) 241. 热滚筒243在图IC中有示出并且在下文中进行更完整地描述。 Heat roll 243 there is shown in FIG IC and be more fully described hereinafter. 在一些实施例中,提供了在微流体器件和HIS/滚筒之间的硬接触。 In some embodiments, there is provided a hard contact between the microfluidic device and HIS / drum. 此外,如下文更详细地描述,在一些实施例中使用真空技术,使微流体器件相对于托架定位并固定住。 Furthermore, as described in more detail using a vacuum technique, in some embodiments, the microfluidic device relative to the bracket is positioned and fixed.

[0123] 通常,运些器件被置于热循环板上,W便使该器件热循环。 [0123] Typically, these devices are placed in transport thermal cycling plate, W will make the device heat cycle. 各种运样的板都可W从下列公司买到:ThermoHybaid Px2 (Rranklin, MA) ;MJ ResearchPTC-SOO (South San Rrancisco, CA) ;E;ppendorf Part組5331 (Wes忧ury,NY) ;Techne Pa;rt#205330 (Princeton, NJ)〇 Various kind of plates can be transported from the following companies buy W: ThermoHybaid Px2 (Rranklin, MA); MJ ResearchPTC-SOO (South San Rrancisco, CA); E; ppendorf Part group 5331 (Wes worry ury, NY); Techne Pa; rt # 205330 (Princeton, NJ) square

[0124] 在一些实施例中,微流体器件与热控制器件接触,使得热控制器件与热控制源热连通,所W作为热控制源的溫度变化的结果,至少一个反应室中的反应溫度会发生变化。 [0124] In some embodiments, the microfluidic devices and thermal control device contacts, so that the heat control devices and thermal control source in thermal communication, as a result of temperature change of the thermal control of the source of the W, at least one reaction chamber reaction temperature change. 在不同的实施例中,热转移器件可W包括半导体(比如娃),可W包括反射材料,和/或可W包括金属。 In various embodiments, the heat transfer device may include a semiconductor W (eg baby), W may include a reflective material, and / or W comprise a metal.

[01巧]热控制器件可W向热转移器件施加力,W便朝着热控制源推动热转移器件。 [01 Qiao] W thermal control device can apply a force to the heat transfer device, W will be moving in the thermal control source to promote heat transfer device. 在不同的实施例中,运种力可W包括机械压力、磁力、静电力、或真空力。 In various embodiments, the force at the W transported species include mechanical pressure, magnetic, electrostatic, or vacuum force. 例如,在一个实施例中, 运种力包括真空力,真空力是通过热控制器件或热转移器件的表面中所形成的通道朝着热转移器件施加的。 For example, in one embodiment, the force comprises a vacuum force kind of operation, a vacuum force through the surface of the device or the hot heat transfer channel devices formed in the heat transfer towards the control applied to the device. 可W检测热控制器件的表面和热转移器件的表面(或表面的一部分)之间所实现的真空度。 W can be detected between the surface of the thermal control device (or a part of the surface) and the surface of the heat transfer device to achieve a degree of vacuum. 运种检测可W用真空度检测器(它位于沿通道远离真空源的一位置处)来执行。 (A position where it is located along the path away from the vacuum source) can be transported species detected W vacuum degree detector to perform. 当真空没有超过预设的水平时,可能出现警报,或者可能进行重新对准。 When the vacuum does not exceed a preset level, an alert may appear, or may be re-aligned.

[01%] 通过使用一个或多个机械或电机械定位器件,阵列器件可W与热控制器件接触。 [01%] by using one or more mechanical or electro-mechanical positioning device, and an array of devices W thermal control device contacts. 该方法的执行过程可W受到自动地控制和监控。 During the execution of the method may be automatically controlled by the W and monitoring. 例如,用自动控制系统可W执行运种自动控制和监控,该自动控制系统与机器人控制系统相连,W便将该阵列器件引入热控制器件或从热控制器件中取出该阵列器件。 For example, the automatic control system may perform operation W kinds of automatic control and monitoring, the automatic control system and robot control system is connected, then the W array device introduced thermal control device or remove the device from the array of thermal control device. 各种反应的进程也可W受到监控。 Process various reactions may be W is monitored. 阳127] 可W提供一种包括热控制器件的单元。 Yang 127] W can provide a thermal control device comprises a unit. 可W提供一种包括阵列器件和热控制器件的系统。 W can provide a device comprises an array of devices and thermal control systems. 为了确保热循环步骤的精确度,在某些器件中,包括传感器是很有用的,运些传感器可用于检测该器件各个区域的溫度。 In order to ensure the accuracy of thermal cycling steps, in certain devices, including sensor is useful, it may be used to transport these sensors detect the temperature of various regions of the device. 一种用于检测溫度的结构是热电偶。 Structure for detecting temperature is a thermocouple. 运种热电偶可W作为底层基板材料上图形化的薄膜导线而产生,或者作为被直接引入微制造的弹性材料中的导线而产生。 Thermocouple can be shipped as a W on the underlying substrate material patterned thin film conductors to produce, or be introduced directly into a microfabricated elastomeric material wire is generated.

[0128] 通过电阻变化,也可W检测溫度。 [0128] The change in resistance, but also at the W detected temperature. 例如,使用常规技术在底层半导体基板上制造的热敏电阻的阻抗变化可W被校准到给定的溫度变化。 For example, using conventional techniques thermistor on the underlying semiconductor substrate made of W impedance changes can be calibrated to a given change in temperature. 或者,热敏电阻可W被直接插入微制造的弹性材料中。 Alternatively, the thermistors W elastic material is inserted directly in microfabrication. 在Wu等人的文章"MEMS Flow Sensors化r Nano-fluidic Applications"(Sensors and Actuators A 89152-158, 2001 年)中,描述了另一种利用阻抗来检测溫度的方法,该文章整体引用在此作为参考。 Wu et al., In the article "MEMS Flow Sensors of r Nano-fluidic Applications" (Sensors and Actuators A 89152-158, 2001 Nian), there is described another impedance to detect the temperature of the use of the method, the reference in this article Reference. 该文章描述了将渗杂多晶娃结构应用于控制和检测溫度。 This article describes the baby doped polycrystalline structure is applied to the control and the detected temperature. 对于多晶娃和其它半导体材料,阻抗的溫度系数可W由杂质的类别和量来精确地控制,由此针对给定的应用使传感器的性能最佳化。 For polycrystalline baby and other semiconductor materials, impedance temperature coefficient W can be accurately controlled by the type and amount of impurities, whereby a given application for the sensor to optimize performance.

[01巧]热-色材料(Thermo-C虹omatic material)是另一种可用于检测扩增器件各区域溫度的结构。 [01 Qiao] hot - colored material (Thermo-C Hong omatic material) is another structure may be used to detect amplification device temperature of each region. 具体来讲,某些材料在经历不同溫度时会急剧地且可复现地改变颜色。 Specifically, certain materials when undergoing different temperatures will rapidly and reproducibly change color. 运种材料可W被添加到溶液中,因为它们历经不同的溫度。 W may be transported kinds of materials added to the solution, as they are after a different temperature. 热-色材料可W形成于底层基板上, 或者被包括到弹性材料中。 THERMAL - W color material may be formed on the underlying substrate, or be included in the elastomeric material. 或者,热-色材料可WW颗粒的形式被添加到样品溶液中。 Alternatively, the heat - colored material can be in the form of WW particles is added to the sample solution. [0130] 另一种检测溫度的方法是使用红外照相机。 [0130] Another method is to use the detected temperature of the infrared camera. 红外照相机和显微镜相结合可W被用来确定整个扩增结构的溫度分布。 Infrared camera and microscope combination W can be used to determine the temperature distribution of the entire amplification structure. 对于合适波长的福射(例如,热福射、红外福射等)而言, 弹性材料的渗透性将有利于运种分析。 For the blessing of the appropriate wavelength emission (for example, heat radiation, to Fu infrared shooting, etc.), the permeability of the elastic material will facilitate transport species analysis. 阳131] 溫度检测的另一种方法是使用热电传感器。 Yang 131] Another method is to use the temperature detection pyroelectric sensor. 具体来讲,一些结晶材料特别是那些呈现出压电行为的材料可W呈现出热电效应。 Specifically, some of the crystalline material, especially those showing the behavior of piezoelectric material W exhibit the thermoelectric effect. 该效应描述了一种现象,即材料晶格的极化和材料上的电压高度依赖于溫度。 This effect describes a phenomenon that the polarization voltage on the material and the material lattice is highly dependent on temperature. 运种材料可W被包括到基板或弹性体中并且被用于检测溫度。 W transported kinds of materials may be included in the substrate or elastomer and is used to detect temperature. 根据本发明的各实施例,其它电学现象(比如电容和电感)都可W被用于检测溫度。 According to various embodiments of the present invention, other electrical phenomena (such as capacitance and inductance) can be used to detect the temperature of the W. 本领域的技术人员将会认识到许多变体、修改和替换。 Those skilled in the art will recognize that many variations, modifications and substitutions.

[0132] 在一个实施例中,成像系统200按下面的方式来工作。 [0132] In one embodiment, the imaging system 200 to work in the following manner. 首先,微流体器件205被牢固地置于托架207上。 First, microfluidic device 205 is stably positioned on the tray 207. 基于微流体器件205的固定的特征(例如,微流体器件的底座的边缘),计算机270使X、y驱动(未示出)移动托架207, W便对准第一X、y位置中的微流体器件。 Based on a fixed feature microfluidic device 205 (e.g., an edge of the base of the microfluidic device), the computer 270 so that X, y drive (not shown) to move the carriage 207, W will be aligned with the first X, y positions microfluidic device. 在一些实施例中,在对准和定位过程中,使用一个或多个基准标记。 In some embodiments, the alignment and the positioning process, or the use of a plurality of reference marks. 在特定的实施例中,系统用户用成像系统来记录一个或多个基准标记的精确坐标。 In certain embodiments, the system user by an imaging system to record the precise coordinates of one or more reference marks. 在其它实施例中,该过程是自动执行的,因为通过定位对称的XY基准物体并除去任何非对称成分便可W精确地计算基准的中屯、。 In other embodiments, the process is performed automatically, as by positioning symmetrical XY reference object and remove any non-symmetrical component W can be accurately calculated on the basis of the village. 在一些实施例中,在对准过程中使用了基准的特征(比如边和角)。 In some embodiments, the alignment process using a reference characteristic (such as edges and corners). 本领域的技术人员将会认识到许多变体、修改和替换。 Those skilled in the art will recognize that many variations, modifications and substitutions.

[0133] 在计算机270的控制下,调节托架207使其定位于光学元件210和212的焦平面中,或者调节光学元件210和212使光学元件210和212的焦平面定位到托架207。 [0133] Under control of the computer 270, the carriage 207 so positioned adjust the focal plane of the optical element 210 and 212, or adjusting optical element 210 and 212 so that the focal plane of the optical element 210 and 212 is positioned to the carriage 207. 在较佳实施例中,视场可W包住整个微流体器件,其中包括微流体器件上存在的许多反应室。 In the preferred embodiment, the field can wrap the entire W microfluidic devices, including many of the reaction chamber present on the microfluidic device.

[0134] 微流体器件的反应室中正发生的化学过程所发出的巧光、化学发光或光信号均被第一透镜系统210收集。 [0134] clever light, chemiluminescence or optical signals chemical reaction process chamber microfluidic device is occurring is issued by the first lens system 210 are collected. 在本发明的一些实施例中,第一透镜系统是多元件光学系统,其中包括一个或多个透镜W及一个或多个孔径。 In some embodiments of the present invention, the first lens system is a multi-element optical system, which includes one or more lenses or W, and a plurality of apertures. 如图2A所示,第一透镜系统210包括多个单独的透镜元件W及成对的透镜元件等。 , The first lens system in FIG. 2A 210 comprises a plurality of individual lens elements W, and the pair of lens elements and the like. 选择第一透镜系统210的光学特性(其中包括焦距f/# 等)W提供期望的光学性能。 Selecting a first lens system 210 of the optical characteristics (including the focal distance f / #, etc.) W provides the desired optical properties. 本领域的技术人员将会认识到许多变体、修改和替换。 Those skilled in the art will recognize that many variations, modifications and substitutions. 图IA 中示出了发射快口215,用于阻挡离光轴预定距离W外的光线,当然本发明对此不作要求。 Figure IA shows the emission fast port 215 for blocking a predetermined distance from the optical axis W light outside, of course, this is not a requirement of the present invention.

[0135] 再次参照图1A,提供了滤光器件213,作为光学组件的一部分。 [0135] Referring again to Figure 1A, the filter element 213 is provided, as a part of the optical assembly. 在一些实施例中, 滤光器件是一种包括许多光学元件的滤光轮213,用于使巧光标记试剂或化学发光标记试剂所产生的巧光发射或化学发光发射通过并且对其进行光学处理。 In some embodiments, the filter device is a filter wheel comprising a number of optical elements 213 for causing light clever labeled reagent or chemiluminescent labeling reagents generated clever light emission or chemiluminescence emission and by its optical deal with. 作为一个示例,在一个实施例中,该发射滤光轮的第一部分适用于使第一巧光染料切™3所产生的巧光发射通过, 该第一巧光染料可W从Amersham Biosciences公司(该公司是新泽西州Piscataway市GE Healthcare的一部分)购买。 As an example, in one embodiment, the first part of the emission filter wheel is applied to the first dye-cut ™ Qiao Qiao light emitted by the light generated by the 3, the first light clever dye W from Amersham Biosciences Inc. ( the company is part of GE Healthcare, Piscataway, NJ City of) purchase. 上述发射滤光轮的第二部分适用于使第二巧光染料切"5所产生的巧光发射通过,该第二巧光染料也可W从Amersham Biosciences公司购买。当然,运些巧光染料的使用不是本发明所要求的。在可选实施例中,使用一种可W从加州Carlsbad 市的Invitrogen公司买到的Alexa巧光染料。作为一个示例,在另一个实施例中,上述发射滤光轮的第一部分适用于使第=巧光染料Alexa Fluor 350所产生的巧光发射通过,该第S巧光染料可W从Invitrogen公司购买。上述发射滤光轮的第二部分适用于使第四巧光染料Alexa Fluor 488所产生的巧光发射通过,该第四巧光染料也可W从Invitrogen公司购买。在下文中将提供与该发射滤光轮有关的其它细节。 The second portion of the emission filter wheel is applied to the second optical dye clever cut "a clever light-emitting 5 produced by the second light clever dye may also be purchased from Amersham Biosciences W company. Of course, some clever optical transport dye the use of the present invention is not required. in an alternative embodiment, the use of a W available from Invitrogen, Carlsbad, Calif available Alexa clever optical dyes. as an example, in another embodiment, the above-described emission filter, Nimbus first portion adapted to cause the first light = Qiao Qiao light-emitting dye Alexa Fluor 350 generated by the first s W clever optical dye may be purchased from Invitrogen Corporation. the second portion of the emission filter wheel is adapted to cause the first four Qiao Qiao light dye Alexa Fluor 488 light emission generated by the fourth clever optical dye may also be purchased from Invitrogen W company. provide other details relating to the emission filter wheel hereinafter.

[0136] 在一些实施例中,滤光器件213和发射快口215都位于第一透镜系统和第二透镜系统之间。 [0136] In some embodiments, the filter device 213 and transmit ports 215 are located between the fast lens system of the first and second lens systems. 在运些实施例中,穿过滤光器件的光线W相对于光轴很小的角度进行传播。 In some embodiments, operation, light passes through the optical device with respect to the optical axis W of the small angle of spread. 对于本领域的技术人员而言,很明显,在光线入射角很小的各区域中的光谱滤光片(例如,干设滤光片)更易于设计,并且有可能提供更窄的总光谱带宽,通过运种窄光谱带宽,各种特征和/或滤光片定位都是本发明所要求的。 For the skilled person that, obviously, the angle of incidence of light in the regions of small spectral filter (e.g., filter set dry) is easier to design, and it is possible to provide total spectral bandwidth is narrower by Yun kind of narrow spectral bandwidth, various features and / or filter of the present invention, the positioning is required. 如图IA所示,滤光器件和发射快口都禪合到计算机270,从而为运些元件提供了计算机控制。 As shown in Figure IA, the filter device and launch fast Zen ports are bonded to the computer 270, so as to transport these elements provide computer control. 此外,对于本领域的技术人员而言,很明显, 可W在光路中设置多个滤光片且很可能是多个完全一样的滤光片,W增大对激发波长的阻隔。 In addition, those skilled in the art, it is quite obvious, the W plurality of filters in the optical path, and is likely to be more of exactly the same filter, W increases the excitation wavelength of the barrier. 在一些实施例中,运些滤光片相对于光轴有一定角度,所W被滤光片反射掉的光线会离开光路。 In some embodiments, these filter operation with respect to the optical axis at a predetermined angle, the filter W are reflected off the light will leave the optical path.

[0137] 在其它实施方式中,某些嵌入染料在结合于双链DNA后会产生剧烈的巧光增强和/或显示出对双链DNA具有强烈的化学亲和力,可用于检测双链扩增DNA。 [0137] In other embodiments, some embedded dye binding to double-stranded DNA in the post will have a dramatic clever optical enhancement and / or showing double-stranded DNA has a strong chemical affinity, can be used to detect amplification of double-stranded DNA . 合适的染料示例包括但不限于SYBR™和Pico Green(俄勒冈州Elugene市的Molecular Probes公司)、漠化乙锭、舰化丙锭、色霉素、日丫晚澄、化echs口3258、Toto-U化yo-1和DAPI (4', 6-二脉基-2-苯基吗I噪盐酸盐)。 Suitable examples include, but are not limited to, dye SYBR ™ and Pico Green (Oregon Elugene City, Molecular Probes Inc.), ethidium desertification, propidium ship, color neomycin, date night Ah Cheng, of echs port 3258, Toto- U of yo-1 and DAPI (4 ', 6- phenyl-2-second pulse noise do I hydrochloride). 关于嵌入染料的使用,在Zhu等人Anal.化em. 66 : 1941-1948(1994)中进行了另外的讨论,该文章整体引用在此作为参考。 Using about embedding the dye in Zhu et al., Anal of em 66:. (1994) 1941-1948 was further discussion, the article entirety incorporated herein by reference. 阳13引图IA也示出了第二透镜系统212。 Yang cited 13 Figure IA also shows a second lens system 212. 穿过滤光器件213和发射快口215的巧光或化学发光发射被第二透镜系统聚焦到检测器260。 Pass through the filter element 213 and the light-emitting fast mouth clever or chemiluminescent emission 215 second lens system is focused to the detector 260. 在一个实施例中,该检测器是CCD照相机阵列,但本发明对此不作要求。 In one embodiment, the detector is a CCD camera arrays, but the present invention this is not required. 在特定的实施例中,使用了一种阵列检测器,大小和微流体器件差不多。 In a particular embodiment, the use of an array of detectors, and microfluidic device similar size. 较佳地,选择检测器阵列260的像素大小,W提供比微流体器件的反应室面积要小的面积,由此使每个反应室对应多个检测器像素。 Preferably, the detector array 260 to select the pixel size, W provides the reaction chamber of the microfluidic device the area ratio of the smaller area, whereby each reaction chamber corresponding to the plurality of detector pixel. 在特定的实施例中,检测器260是一种具有大约15ymxl5iim像素的CCD阵列。 In a particular embodiment, the detector 260 is a CCD having about 15ymxl5iim pixel array.

[0139] 对于本文所提供的微流体器件,可W使用许多不同的检测方案。 [0139] For microfluidic devices provided herein, W can be detected using a number of different programs. 合适系统的选择部分程度上基于所检测的事件和/或试剂的类型。 And / or type of suitable reagent system of choice on the part of the degree based on the detected event. 运些检测器可W被设计成检测许多不同的信号类型,包括但不限于来自下列物质的信号:放射性同位素、巧光团、生色团、电子密集颗粒、磁性粒子、自旋标记物、化学发光分子、电化学活性分子、酶、辅因子、连接于核酸探针的酶,W及酶底物。 These detectors can be transported W is designed to detect a number of different signal types, including but not limited to a signal from the following materials: radioisotopes, clever light, chromophores, electron dense particles, magnetic particles, spin labels, chemical luminescent molecules, electrochemically active molecules, enzymes, cofactors, enzymes linked to nucleic acid probes, W, and an enzyme substrate.

[0140] 适于本发明微流体器件使用的说明性的检测方法包括但不限于:光散射、多通道巧光检测、UV和可见光波长吸收、发光、示差反射、和激光共聚焦扫描。 [0140] Illustrative detection methods suitable for the microfluidic device of the present invention include, but are not limited to: light scattering, light clever multi-channel detection, UV and visible wavelength absorption, luminescence, differential reflection, and confocal laser scanning. 可用于某些应用中的其它检测方法包括:闪烁亲近测定技术、放射化学检测、巧光偏振各向异性、巧光寿命、巧光相关频谱学(FCS)、时间分辨能量转移(TRET)、巧光共振能量转移(FRET)、W及像生物发光共振能量转移度RET)运样的变化形式。 Can be used in some applications other detection method comprising: scintillation proximity assay techniques, radiochemical detection, polarization anisotropy clever, clever light life, clever optical spectrum related science (FCS), time-resolved energy transfer (TRET), Qiao optical resonance energy transfer (FRET), W and the like bioluminescence resonance energy transfer of RET) kind of transport in the form of change. 其它检测选择包括电阻、电阻率、阻抗和电压的检测。 Other options include detecting resistance, resistivity, impedance, and voltage detection. 阳141] 在一些实施例中,在"检测部分"或"检测区域"进行检测。 Yang 141] In some embodiments, the "test section" or "detection zone" for testing. 运些术语和其它相关术语是指微流体器件上进行检测的那部分。 Transportation and other related terms These terms refer to that part of detecting the microfluidic device. 在一些微流体器件中,检测部分常常是微流体器件中存在的反应室。 In some microfluidic devices, often detecting portion microfluidic device present in the reaction chamber. 其它器件的检测部分可W在与交叉点相邻的流体通道区域内、交点本身、或包括交点及其周围区域的区域。 Detecting at the W part of other devices in the point of intersection of adjacent fluid channel region, the intersection itself, or a region including the intersection of its surrounding area. 阳142] 检测部分可W连接到一个或多个显微镜、二极管、光激励器件(例如,激光器)、光电倍增管、处理器、W及上述的组合,它们配合在一起来检测和特定事件和/或试剂相关联的信号。 Male 142] detecting portion may be connected to one or more of W microscopes, diodes, light excitation devices (e.g., lasers), photomultiplier tubes, processors, and combinations of the above photoelectric W, which fit together to detect specific events and / or agents associated with the signals. 通常,待测信号是光信号,由一个或多个光检测器在检测部分中对其进行检测。 Typically, the signal under test is an optical signal, by one or more light detectors detect it in the detection portion. 光检测器可W包括一个或多个光电二极管(例如,雪崩光电二极管)、引向如光电倍增管的光纤光导、显微镜、和/或视频照相机(例如,CCD照相机)。 W photodetector may comprise one or more photodiodes (e.g., avalanche photodiodes), toward the light guide fiber as the photomultiplier tube, a microscope, and / or a video camera (e.g., the CCD camera). 阳143] 检测器可W在微流体器件内被微制造,或者可W是分离的元件。 Yang 143] W in the detector may be microfabricated microfluidic device, or may be W is an element separation. 如果检测器作为分离的元件存在并且微流体器件包括多个检测部分,则在任何给定的时刻可W在单个检测部分内进行检测。 If the detector exists as a separate element and the microfluidic device comprises a plurality of detecting portions, at any given moment W can be detected in a single detection section. 作为一个具体的示例,微流体器件可W被附在可平移的工作台上并且在显微镜物镜下进行扫描。 As a specific example, the microfluidic device W can be attached to the translating table and scanned under a microscope objective. 如此获得的信号接下来传递到处理器,W便于信号解释和处理。 Signal thus obtained is transmitted to the next processor, W easy to interpret and process the signal. 也可W使用光电倍增管的阵列。 W can also use an array of photomultiplier tubes. 另外,可W使用运样的光学系统,它们能够从所有不同的检测部分中同时收集信号,而且能够判定来自每一个部分的信号。 Further, the sample used at the W optical transport system, which can collect signals from all the different detection sections simultaneously, and can be determined from the signal of each portion.

[0144] 外部检测器是可W使用的,因为所提供的器件完全或大部分都由对受监控的波长透明的材料制成。 [0144] W external detector is used, as provided by the device is made up mostly or entirely of a material transparent to the wavelength monitored is made. 该特征能够使本发明的器件使用许多光学检测系统,运对于常规的娃基微流体器件而言是不可能的。 This feature enables the device of the present invention uses many optical detection system, for the transport of conventional microfluidic device in terms of baby group is not possible.

[0145] 本发明的特定实施例使用了形如CCD照相机的检测器W及可提供大视场和高数值孔径的光路,W使从每一个反应室中收集到的光量最大化,由此增大了检测灵敏度。 [0145], to maximize the amount of light W so collected from each of the reaction chamber to a particular embodiment of the invention embodiment uses a shape such as a CCD camera detector W and can provide a large field of view and a high numerical aperture of the optical path, thereby increasing Great detection sensitivity. 在本实施例中,该CCD被用作光电检测器的阵列,其中每一个像素或像素组对应于一个反应室, 而非被用来产生该阵列的图像。 In the present embodiment, the CCD is used as the photodetector array, wherein each pixel or group of pixels corresponds to a reaction chamber, instead of being used to generate an image of the array. 因此,可W设计或改变光学元件,使得在检测器处图像质量下降或图像变得模糊,W增大该光学系统的可用场深,从而从每一个反应室中收集更多的光线。 Thus, W design or alter the optical element, such that at the detector drop in image quality or image blurring, W to increase the depth of field of the optical system can be used to collect more light from each of the reaction chamber. 特别是,因为本发明的一些实施例中所预期的试验包括使用巧光染料的生物试验,运些染料因暴露于激发光线而使光反差变小(photobleach),因此限制了从给定样品中可W 获得的信号光子的总数,有限信号光子的有效收集在仪器中是很重要的。 In particular, because of some embodiments of the present invention is expected to test include the use of biological tests clever optical dyes, dye shipped some exposure to the excitation light leaving the optical contrast becomes smaller (photobleach), thus limiting from a given sample the total number of W can get a signal photon and effectively collect a limited signal photons in the instrument is very important. 对于任何光学系统,化en化e的种种考虑设及物像NA(数值孔径)和总的系统放大率捆为像方NA通常是有限的(例如,对高入射角光线而言在CCD表面上有反射损耗),对于任意高的放大率而言无法同时实现任意高的物方(样品那边)NA。 For any optical system, based en the various considerations set of e transitive like NA (numerical aperture) and total system magnification of the image side NA bundle is usually a limited (for example, high angle of incidence of light in terms of the CCD surface reflection loss), it can not simultaneously achieve a high magnification for any arbitrarily high in terms of the physical side (sample side) NA. 事实上,更大的系统放大率可W允许更高的物方NA,同时不要求像方NA同时上升(该上升有可能有害,原因如上)。 In fact, the greater the magnification system may allow higher W NA object side, while at the same time does not require the image side NA rise (the rise has potentially harmful reasons as above). 结果,在所描述的系统中,大CCD(例如,30. 7mmx30. 7mm)焦平面阵列被用于允许1 : 1的光学系统(即系统放大率为1)。 As a result, in the described system, the CCD large (.. E.g., 30 7mmx30 7mm) focal plane array is used to allow 1: 1 of the optical system (i.e., a magnification system). 运允许选取NA为0. 36,同时CCD上的像方NA为0. 36,运便提供了相对于表面反射损耗的合理的性能。 Transported to allow selection of NA 0.36, and on the image side NA CCD is 0.36, shipped it provides reasonable performance with respect to surface reflection losses. 阳146] 在一些实施例中,更大的物方NA产生减小的物方焦深,因此对于样品中给定的反应室深度,检测器处的模糊更严重了(假定因焦深引起的模糊大于或等于因透镜像差和其它问题所引起的模糊),从而如果在反应室之间实现了低信号串扰则限制了样品处各反应室之间可允许的最小间隔。 Yang 146] In some embodiments, the bigger the object side NA produce a reduced depth of focus on the object side, so for a given sample of the reaction chamber depth, blur at the detector is more serious (assuming that the depth of focus caused by Fuzzy or greater because the lens aberration and other problems caused by the blur), so if between the reaction chamber is to achieve low crosstalk limits the minimum interval between each sample point allows the reaction chamber. 在与1 : 1光学系统相结合的情况下,如果想要避免其表面处的反射损耗,则运种物方NA考虑与CCD (或娃检测器)上一般期望的~0. 5NA最大值很一致。 With 1: 1 case of combined optical system, if you want to avoid reflection losses at the surface thereof, the species transported object side NA consider CCD (or baby detectors) general desired maximum value is ~ 0 5NA consistent. 该1 : 1成像透镜系统还固有地免受大多数奇数阶像差的影响,从而增大了该特定放大率(M= 1)的优势。 The 1: 1 imaging lens system is also inherently immune to most of the odd-order aberrations, thereby increasing the specific magnification (M = 1) advantage. 作为详细的系统的设计,在本发明的一些实施例中,使用了具有检测器的I : I光学系统,它和待成像的微流体系统一样大或更大。 As the detailed design of the system, in some embodiments of the present invention used with a detector I: I optical system, and it is as large or larger to be imaged microfluidic system. 阳147] 在其它实施例中,可能存在和检测器(例如,CCD焦平面阵列)尺寸有关的成本限审ij。 Yang 147] In other embodiments, there may be and the detector (for example, CCD focal plane array) size limit costs associated with the trial ij. 例如,目前一些高量子效率、全画面CCD阵列具有27. 6mmx27. 6mm的尺寸。 For example, the current number of high quantum efficiency, full-frame CCD array with 27. 6mmx27. 6mm size. 和尺寸为30. 7mmx30. 7mm的微流体器件相比,该检测器稍微小一点,从而产生0. 88的系统放大率,作为用于上述系统的一种设计。 And size 30. 7mmx30. 7mm compared microfluidic device, the detector is slightly smaller, resulting in a 0.88 magnification system, as a design for the system described above. 因为接近系统放大率M= 1,所W针对运种放大率,与上述检测器(像方)入射NA有关的种种约束条件均得到满足。 Because the system is close to the magnification M = 1, the W species transported for magnification, and said detector (image side) to various constraints related to the incident NA have been satisfied.

[0148] 在其它实施例中,可W指定各反应室的给定XY平面(和光轴相垂直)间隔和大小(例如,在XY平面中实现期望的样品-反应室密度),同时反应室的最小总体积仍然受到约束(例如,实现最小所需化学体积,W避免因试剂或目标分子数目太小而导致的过大的统计波动,或简单地实现所需的最少数目的巧光或其它发光分子或物体)。 [0148] In other embodiments, at the W specify a given XY planes of the reaction chamber (optical axis perpendicular) spacing and size (e.g., to achieve a desired in the XY plane of the sample - the reaction chamber density), while the reaction chamber the minimum total volume remains restrained (for example, to achieve the minimum desired chemical volume, W to avoid excessive statistical fluctuations due to the number of agents or the target molecule is too small to cause or simply to achieve the required minimum number of clever light or other light emitting molecules or objects). 在运种情况下,可能有必要平行于Z轴(光轴)来延伸反应室,使得每一个反应室的总体积仍然等于或大于一些最小数字。 In case the operation, it may be necessary parallel to the Z axis (optical axis) extending to the reaction chamber such that the total volume of each reaction chamber is still greater than or equal to the minimum number of figures. 假定焦深所引起的模糊大于或等于由透镜像差和其它问题所导致的模糊, 考虑到焦深,对于给定的物方NA而言,沿Z轴作更大的延伸(从而产生高纵横比或柱状反应室,运使待询问的样品沿Z轴集中)通常将使检测器处的反应腔图像更模糊。 Fuzzy assumed depth of focus caused by the lens is greater than or equal to the fuzzy aberration and other problems caused, taking into account the depth of focus for a given object side NA is concerned, along the Z axis for greater extension (resulting in high aspect ratio or cylindrical reaction chamber, making transport a sample to be asked along the Z axis focus) will usually a reaction chamber at the image detector more ambiguous. 在一些情况下,运将导致使用更低的物方NA。 In some cases, the use of transport will lead to a lower object side NA. 使用更低的NA透镜系统可W允许焦深更大,因此,从平行于光轴延伸的反应室中更好地进行光线收集,同时通常不会引起相邻或相近的反应室之间的光信号串扰。 Using a lower NA of the lens system at the W allows a greater depth of focus, therefore, the optical axis extending parallel to the reaction chamber for better light while collecting the light does not usually cause adjacent or close to the reaction chamber between the signal crosstalk. 运样,可W使XY平面(存在于光轴的地方)中反应室的密度更大,却没有过度的串扰,同时通过在Z方向(平行于光轴)延伸,使总的反应室体积保持很大。 Like transportation, can make the XY plane W (where present in the optical axis) greater density in the reaction chamber, but without excessive crosstalk, through the Z direction (parallel to the optical axis) extends, bringing the total volume of the reaction chamber maintained great. 在运种情况下,或者在更低的物方NA尚可接受的情况下(例如,情况一,各反应室之间更大的XY 间隔使检测器处的反应腔-图像更模糊,同时却没有过多的串扰;情况二,在非光发射应用中,更高的NA不是必需的;情况S,有足够多的样品W至于光白化不是问题;情况四,没有光白化的样品,像降低可接受的系统灵敏度运样的情形),较低的系统放大率(M< 1)可能是合适的,尤其是当M > NAyo. 5时,M > NAyo. 36更佳,其中NA。 In the transport case or, in the case of a lower object-side NA is acceptable (for example, the case of a larger XY spacing between the reaction chamber so that the reaction chamber at the detector - image is more blurred, but at the same time not too much crosstalk; case 2, the non-light-emitting applications, the higher the NA is not required; the case of S, there are a sufficient number of samples W as for whitening light is not a problem; case 4, there is no light whitened samples, like reduced acceptable kind of transport system sensitivity case), low magnification system (M <1) may be appropriate, particularly when M> NAyo. 5 Shi, M> NAyo. 36 is better, where NA. =物方NA,或者M > NA。 = Object side NA, or M> NA. / NAdet,其中NAdet等于可允许到检测器表面上的最大NA,同时对于检测器而言没有过大的反射/插入损耗(对于典型的CCD而言,NAdet= 0. 36~0. 5)。 / NAdet, where NAdet equal to the maximum allowable NA on the detector surface, and for the purposes of the detector without excessive reflection / insertion loss (for a typical CCD is concerned, NAdet = 0. 36 ~ 0. 5).

[0149] 当物方焦深和/或模糊要求并不需要物方NA《0. 36或0. 5或者更一般地讲酷。 [0149] When the object-side depth of focus and / or ambiguous requirements do not need the object side NA "0. 36 or 0.5 or more generally cool. 《NAdet时,基于化emlue的种种考虑(如上所述)期望用更大的检测器,因为对于给定的NA。 "NAdet when, based on the considerations of emlue (as described above) with the desired larger detector, because for a given NA. ,更大的M(对于给定的样品尺寸,通常需要更大的检测器)将允许更小的NAi (像方NA)。 Larger M (for a given sample size usually requires a larger detector) will allow smaller NAi (the image side NA). 因此,当光收集要求(例如,实现某些试验灵敏度)需要很大的NA。 Thus, when the light collection requirements (for example, to achieve certain test sensitivity) requires a lot of NA. (由酷。> NA det来定义)并且其它设计考虑(例如,成本)允许很大的时NA。 (Defined by cool.> NA det) and other design considerations (for example, cost) allows a great time NA. ,期望M更大,使得检测器处的损耗最小化。 Expect more M such that the loss at the detector is minimized. 在运种实施例中,有用的是,使用一个或多个CCD器件等检测器件,其尺寸要大于待成像的微流体器件的面积。 In such an embodiment of the transport, it is useful to use one or more CCD devices and other detection devices, its size is greater than the microfluidic device to be imaged area. 使用运种大检测器可W增大光学系统的放大率,因此(通过化emlue的种种考虑),对于到检测器的固定入射NA,从样品中收集光线的NA可W更高(例如,通过CCD表面处更高入射角的入射光的反射损耗来实现后者) W can be transported using an NA higher seed large detector can increase the magnification of the optical system W, therefore, to the detector fixed incident NA, to collect light from the sample (by considerations of emlue a) (for example, by CCD surface reflection losses at higher angles of incidence of the incident light to achieve the latter)

[0150] -种较佳的检测器使用CCD照相机和可W提供大视场和高数值孔径的光路,从而使从各反应室中收集的光的量最大化,由此增大了检测灵敏度。 [0150] - species preferred detector uses a CCD camera provided at the W and the large field of view and a high numerical aperture of the optical path, so that to maximize the collection of light from each reaction chamber volume, thereby increasing the detection sensitivity. 关于运一点,CCD是作为光电检测器的阵列而被使用的,其中每一个像素或像素组对应于一个反应室,而非被用来产生该阵列的图像。 That on operation, the CCD is used as the photodetector array is used, wherein each pixel or group of pixels corresponds to a reaction chamber, instead of being used to generate an image of the array. 因此,可W改变光学系统,使得图像质量下降或散焦,从而增大光学系统的场深,W便从每一个反应室中收集更多的光线。 Thus, W changing the optical system so that the image quality, or defocus, thereby increasing the depth of field of the optical system, W begins with each reaction chamber to collect more light. 在一些实施例中,有用的是,使用高纵横比或圆柱形反应室,使要由检测器询问的样品沿光学系统的光轴集中,并且较佳地通过使图像散焦来增大场深。 In some embodiments, it is useful, the use of high aspect ratio or cylindrical reaction chamber, so that the optical axis to be focused by the detector asks sample along the optical system, and preferably by defocusing the image to increase the depth of field . 使用低NA透镜系统,较佳地使用两侧对称的透镜系统。 Use low-NA lens system, preferably using a bilaterally symmetrical lens system. 有用的是, 使用一个或多个CCD器件等检测器件,其尺寸大于待成像的微流体器件的面积。 It is useful to use one or more CCD devices and other detection devices, its size is larger than the area to be imaged microfluidic devices. 与低NA光学系统结合使用,可W实现改进的检测灵敏度。 Use in conjunction with the low NA optical system W can achieve improved detection sensitivity. 阳151] -种检测系统可W包括光源,用于激发报道物来产生可检测的信号。 Yang 151] - W detection system may include a kind of a light source for exciting the reporter to produce detectable signal. 所用光源的类型取决于待激发的报道物的性质。 The type used depends on the nature of the light source to be excited by the reporter. 合适的光源包括但不限于激光器、激光二极管、白光源、和高强度灯。 Suitable light sources include, but are not limited to, lasers, laser diodes, white light source, and high-intensity lights. 如果使用激光器,则激光器可W用来在一组检测部分上或单个检测部分上进行扫描。 If using a laser, the laser can be used for W scans on a group or individual detecting portion detecting portion. 激光二极管可W被微制造成微流体器件本身。 W laser diode may be micro fabricated microfluidic device itself. 或者,激光二极管可W被微制造成位于微流体器件附近的另一个器件,它被用来进行热循环反应,使得该二极管的激光被引至检测部分。 Alternatively, laser diodes can be microfabricated W to another device located near the microfluidic device, which is used to carry out a thermal cycling reaction such that the laser diode is directed to the detection section. 阳152] 检测可W包括许多非光学方法。 Yang 152] W detection may include a number of non-optical methods. 例如,检测器可W包括溫度传感器、电导传感器、 电势传感器(例如,pH电极)和/或测量电流的传感器(例如,监测氧化和还原反应)。 For example, W detector may include a temperature sensor, conductivity sensor, a potential sensor (for example, pH electrode) and / or measuring current sensors (for example, to monitor oxidation and reduction reactions). [0153] 某些嵌入染料在结合于双链DNA时会产生剧烈的巧光增强和/或显示出对双链DNA具有强烈的化学亲和力,它们可用于检测双链扩增DNA。 [0153] In some embedded dye binding to double-stranded DNA will produce severe clever optical enhancement and / or showing double-stranded DNA has a strong chemical affinity, which can be used to detect amplification of double-stranded DNA. 合适的染料示例包括但不限于SYBR™和Pico Green (俄勒冈州化gene市的Molecular Probes公司)、漠化乙锭、舰化丙锭、色霉素、日丫晚澄、化echst 33258、Toto-l、化yo-lW及DAPI(4',6-二脉基-2-苯基吗|噪盐酸盐)。 Suitable examples include, but are not limited to, dye SYBR ™ and Pico Green (Oregon City of gene Molecular Probes Inc.), ethidium desertification, propidium ship, color neomycin, date night Ah Cheng, of echst 33258, Toto- l, of the yo-lW and DAPI (4 ', 6- second pulse-2-phenyl it | noise hydrochloride). 关于嵌入染料的使用,在Zhu等人Anal. Chem. 66 :1941-1948 (1994)中进行了另外的讨论,该文章整体引用在此作为参考。 On the use of an intercalating dye, 66 in Zhu et al. Anal Chem:. 1941-1948 (1994) were further discussed in the article reference in its entirety herein by reference. 阳154] 如图IA所示,本发明的一些实施例提供了1 : 1成像系统,用于产生并检测巧光、 化学发光、生物发光W及来自微流体器件的其它信号。 Male 154] As shown in Figure the IA, some embodiments of the present invention provides a 1: 1 imaging system, and for generating a coincidence detection light, chemical, bioluminescence, W, and other signals from the microfluidic device. 在所使用的图像检测器件和待成像的样品一样大的一些实施例中,提供了1 : 1成像系统。 As in the image detection device used and the sample to be imaged large some embodiments, there is provided a 1: 1 imaging system. 通过提供大视场(量级在若干个平方厘米)的1 : 1成像,本发明的各实施例提供了增大的数值孔径(NA)光学系统。 By providing a large field of view (in the order of several square centimeters) of 1: 1 imaging, various embodiments of the present invention provides increased numerical aperture (NA) of the optical system. 因为光收集效率大约正比于NA2,所W本发明的一些实施例所提供的NA增大能够从反应室中收集到合适的巧光信号,运些反应室包括量级在一纳升到数十纳升的反应体积W及量级为1. 0 纳摩尔的活性巧光团浓度。 Since light collection efficiency is approximately proportional to NA2, NA some embodiments of the present invention, there is provided by W can be collected from the reaction chamber to increase the optical signal to an appropriate coincidence, the reaction chamber comprises some luck in the order of several tens of nanoliters to a nanoliter reaction volumes and the order of 1.0 W nanomolar concentrations of active clever light group. 在其它的实施例中,浓度为皮摩尔范围中的活性巧光团提供合适的巧光信号。 In other embodiments, the concentration of the optical signal to provide a suitable clever as clever picomolar range active light group.

[0155] 另外,本发明的各实施例提供了运样的成像系统,它们稍微减小并构成的图像范围从大约和物体大小一样到大约是物体大小的一半。 [0155] In addition, embodiments of the present invention provide a kind of transport imaging systems, and they are slightly reduced range of images and objects constructed from approximately the same size to about half the size of the object. 例如,在一个实施例中,微流体器件的空间区域的图像被传输且被拍摄,该空间区域和不止96个反应室相关联。 For example, in one embodiment, the image of the spatial region of the microfluidic device is transmitted and photographed, and the spatial region than the reaction chamber 96 is associated. 通过使用比微流体器件的空间区域稍小或大致相等的图像检测空间区域,图像检测器件被用于拍摄上述空间区域的图像。 By using the space area smaller than the microfluidic device is substantially equal to or spatial image detection area, the image sensing device is used to capture images in the space area. 仅作为示例,微流体器件的空间区域的面积比上该空间区域的像的面积得到的比例可W是1 : 1、1 : 0. 99、1 : 0. 95、1 : 0. 9、1 : 0. 88、1 : 0. 85、1 : 0. 8、 1 : 0. 7、1 : 0. 6和1 : 0. 5。 Only as an example, the ratio of the area of the spatial region of the microfluidic device of the image area ratio of the spatial region is obtained at the W 1: 1, 1: 0. 99,1: 95,1 0.: 0. 9,1 : 0. 88,1: 85,1 0.5: 0.8, 1: 0. 7,1: 0.6 and 1: 0.5. 在一些实施例中,该比例小于1 : 0. 5。 In some embodiments, the ratio is less than 1: 0.5. 运些特定的比例只是示例性的,因为针对成像系统来选择的比例将取决于特定的应用。 Transport some specific ratio is merely exemplary, as the ratio for the imaging system to choose depends on the specific application. 阳156] 在一些实施例中,光学成像系统包括大约3cmx3cm的视场。 Yang 156] In some embodiments, the optical imaging system includes a field of view of about 3cmx3cm. 在其它实施例中,光学成像系统包括范围从大约Icmxlcm到大约5cmx5cm的视场。 In other embodiments, the optical imaging system includes a range from about Icmxlcm to the field of view of about 5cmx5cm. 在特定的实施例中,提供大小为2cmx2cm、2. 5cmx2. 5cm、2. 76cmx2. 76cm、3. 07cmx3. 07cm、3. 5cmx3. 5cm 和4cmx4cm 的物体视场。 In a particular embodiment, there is provided size 2cmx2cm, 2. 5cmx2. 5cm, 2. 76cmx2. 76cm, 3. 07cmx3. 07cm, 3. 5cmx3. 5cm and field 4cmx4cm objects. 通常,光学成像系统的视场被选择成对应于微流体器件的空间区域(例如,包括许多感兴趣的反应室的区域)。 Typically, the field of view of the optical imaging system is selected to correspond to the spatial region of the microfluidic device (e.g., a reaction chamber including a number of areas of interest). 阳157]此外,本发明的各实施例提供了具有许多数值孔径的光学成像系统。 Male 157] Furthermore, various embodiments of the present invention provides an optical imaging system having a plurality of numerical aperture. 作为一个示例,根据各种实施例,提供了范围从0. 1到0.5的NA。 As one example, in accordance with various embodiments, it provides a range of from 0.1 to 0.5 NA. 在特定的实施例中,提供了大小为0. 15、0. 18、0. 2、0. 23、0. 25、0. 3、0. 36 和0. 4 的多种NA。 In a particular embodiment, there is provided a size 0. 15,0. 18,0. 2,0. 23,0. 25,0. 3,0. 36 and 0.4 of a variety of NA. 阳15引光学成像系统的空间分辨率通常是图像检测器件中像素大小的函数。 Yang cited the optical imaging system 15 is generally a function of the spatial resolution of the image sensing device in pixel size. 在本发明的一些实施例中,放大率(在一些实施例中等于1)和检测器中的像素大小将决定和每一个反应室相关联的像素的数目。 In some embodiments of the present invention, the magnification (equal to 1 in some embodiments) and the detector will determine the pixel size and number of each reaction chamber associated pixel. 通常,每一个反应室最好和多个检测器像素相关联。 Typically, each reaction chamber is preferably a plurality of detectors and associated pixel. 例如,在1 : 1成像系统中,如果反应室一边是45微米,则它将与多达9个方形像素(其边等于15 微米)重叠。 For example, in a 1: 1 imaging system, while if the reaction chamber is 45 m, and it will be up to 9 square pixels (whose sides equal to 15 micrometers) overlap. 因此,根据本发明的各实施例,与每一个反应室相关联的像素数目介于从1到100的范围中。 Thus, in accordance with various embodiments of the present invention, each of the number of pixels with a reaction chamber between the associated range of 1-100. 例如,根据本发明的一些实施例,与每一个反应室相关联的有4个像素区域、 9个像素区域、16个像素区域、25个像素区域、36个像素区域、49个像素区域、64个像素区域和81个像素区域。 For example, according to some embodiments of the present invention, with each reaction chamber associated with four pixel regions, 9-pixel regions, 16 pixel regions, 25 pixel regions, 36 pixel regions, 49 pixel regions, 64 pixel areas and 81 pixel area. 阳159] 在本发明的各实施例中,使用范围介于Iym2~900 ym2之间的像素大小。 Yang 159] In various embodiments of the present invention, the range between the pixel size Iym2 ~ 900 ym2 between. 例如, 在本发明的各种实施例中,使用了边长为1 ym、2 ]im、3 ]im、4 ]im、5 ]im、10 5 ym、 15 ym、20 ym、25 ym、30 ym的正方形像素。 For example, in various embodiments of the present invention, the use of a side length of 1 ym, 2] im, 3] im, 4] im, 5] im, 10 5 ym, 15 ym, 20 ym, 25 ym, 30 ym square pixels. 对于本领域的技术人员而言,很明显,像素大小、 检测器阵列尺寸、和每个阵列的像素数目是彼此相关的。 For the skilled person that, obviously, the number of pixels in the pixel size, the size of the detector array, and each array are related to each other. 在可选的实施例中,使用矩形像素,其像素面积介于1 y m2~900 ym 2之间。 In an alternative embodiment, the use of rectangular pixels, its pixel area is between 1 y m2 ~ 900 ym 2. 阳160] 此外,根据本发明各实施例,使用了检测器阵列(也被称为图像检测器件,其中包括不同范围的像素数)。 Male 160] In addition, according to various embodiments of the present invention, using a detector array (also referred to as an image detection device, which comprises a number of pixels of different ranges). 阵列大小介于512巧12像素区域到3, 000x3, 000像素区域。 Qiao array size between 512 to 12 pixel region 3, 000x3, 000 pixel area. 根据检测器阵列的可获得性,在一些实施例中可W使每个阵列具有更多的像素数目。 According to the availability of the detector array, in some embodiments W may each array having a greater number of pixels. 在特定的实施例中,使用大小为1,024x1,024像素区域和2, 048x2, 048像素区域的阵列。 In a particular embodiment, the array size used 1,024x1,024 pixels and region 2, 048x2, 048 pixel area. 阳161] 本发明的各实施例提供了一种光学成像系统,它具有若干系统参数。 Male 161] Embodiments of the present invention provides an optical imaging system having a plurality of system parameters. 例如,在本发明的各实施例中,可W使用大于35mm的工作距离,例如45. 92mm。 For example, in various embodiments of the present invention may be greater than the working distance of 35mm using the W, for example, 45. 92mm. 在另一个实施例中,提供了平均为13.44 ym的均方根(RM巧光斑直径,其最大值为17.85 ym。此外,在本发明的各实施例中,实现约 5 %的照明变化。在一些实施例中,光学成像系统总长是542. 1mm,其最大滤光片AOI为12. 56度,在滤光片处最大光束直径为76mm,失真< 0. 10%,并且最大透镜直径为5. 512英寸。 阳162] 图IB是本发明一实施例的成像系统的简化图。在一些实施例中,图IB所示的成像系统被用于给微流体器件成像,该微流体器件包括用于执行蛋白质结晶处理的器件。关于图IB所示成像系统及相关联的微流体器件的其它细节在下列文献中可W找到:共同待批且共同拥有的美国专利申请10/902,494(2004年7月28日提交),W及美国专利申请10/851,777 (2004年5月20日提交),运两个专利申请引用在此作为参考。特别是,在运两个专利申请中,可W找到关于本发明各实施例所提供的微流体器件及其和图IB所示成像系统结合使用的其它细节。运些图只是示例,不应该过多限制权利要求书的范围。本领域的技术人员将会认识到许多变体、替换和修改。 In another embodiment, there is provided an average of 13.44 ym rms (RM clever spot diameter, a maximum value of 17.85 ym. In addition, in various embodiments of the present invention, to achieve approximately 5% of the lighting changes. in some embodiments, the total length of the optical imaging system 542. 1mm, the maximum filter AOI of 12.56 degrees, the maximum beam diameter of the filter is 76mm, the distortion <0.10%, and the maximum diameter of the lens 5.512 inches. male 162] Figure IB is a simplified diagram of an image forming system according to an embodiment of the present invention. in some embodiments, the imaging system shown in FIG. IB is used to image a microfluidic device, the microfluidic device comprising the device used to perform protein crystallization process on the imaging system shown in Fig. IB and other details of the microfluidic device associated W can be found in the following documents: co-pending and commonly owned U.S. Patent application 10 / 902,494 (2004 July 28 submitted), W and U.S. Patent application 10 / 851,777 (may 20, 2004 submission), transported two patent applications are incorporated herein by reference. in particular, shipped two patent applications may be W find the range on the microfluidic device and the examples provided and various other embodiments of the present invention is shown in detail IB imaging systems used in conjunction with the op these figures are only examples and should not be too restrictive claims. skill in the art art will recognize many variations, alternatives, and modifications.

[0163] 在一个实施例中,成像系统(10) W下面的方式操作。 [0163] In one embodiment, the imaging system (10) W following manner. 首先,微流体器件(30)牢固地设置在工作台(20)上。 First, microfluidic device (30) fixedly disposed on the table (20). 基于微流体器件(30)的固定特征,计算机(110)使X,y驱动(25)去移动工作台(20),W便使第一X,y位置中的微流体器件(30)与多个基准标记中的第一基准标记对准,其中基准标记被嵌入微流体器件内(离反应室中屯、点的Z维度距离已知),基于该固定特征的推算定位通过成像设备化0)达到聚焦。 Based microfluidic device (30) is fixed characteristic, the computer (110) so that X, y drive (25) to the movable table (20), W will make the first X, y position in the microfluidic device (30) with multiple a reference mark of the first reference mark is aligned, in which the reference mark is embedded within the microfluidic device (Tun from the reaction chamber, Z dimensions from a known point), based on the fixed feature of the imaging device by dead-reckoning 0) achieve focus. 然后,系统用户使精确的基准坐标与像系统重合。 Then, the system user to make an accurate reference coordinate system coincides with the image. 在用户的辅助下,两个或更多个其它的基准标记也同样地被绘制。 In aid of the user, two or more other reference marks are similarly drawn. 在其它实施例中,该过程是自动的,因为通过定位对称的XY基准物并除去任何非对称的成分,便可W精确地计算运些基准的形屯、。 In other embodiments, the process is automatic, because by symmetrical XY positioning reference material and remove any non-symmetrical component W can accurately calculate the transport of these benchmarks shaped village. 成像设备化0)在计算机(110)的控制下接下来调节焦平面(100)的Z维度定位W便聚焦到基准标记。 The image forming apparatus of 0) in the computer (110) Next, adjust the focal plane of the control (100) of the Z dimension W will focus on positioning the reference mark. 例如,一旦聚焦到第一基准标记,成像系统就获得了微流体器件(30)的第一X,y坐标图像,从而寻找成像设备化0)的视场之内其它的基准标记。 For example, when focused to a first reference mark, the imaging system is obtained a microfluidic device (30) in a first X, y coordinates of the image, in order to find the image forming apparatus of 0) within the field of view of the other reference mark. 在较佳实施例中,该视场可W包括整个测定单元。 In the preferred embodiment, the W include the entire field of view can be a measurement unit. 然后,计算机分析该第一X,y坐标图像W确定微流体器件是否具有偏斜和延伸,并且如果确定有偏斜或延伸,贝U 变换该第一x,y图像W使微流体器件的图像和坐标图成为理想的坐标图。 Then, the computer first analyzes the X, y coordinates of the image W is determined whether the microfluidic device has a deflection and extension, if it is determined there is skewed or extending, transforming the first shell U x, y W image to the image of the microfluidic device and plot the ideal plot. 随后在图像减影和掩模步骤中使用该理想的坐标图。 Then use the ideal graph in image subtraction and masking steps.

[0164] 在较佳实施例中,当微流体器件X,y坐标图像与理想的坐标图对准时,系统接下来通过比较X,y坐标图中的基准标记位置和理想的存储图像图的X,y坐标图中的基准标记位置,来确定微流体器件中是否存在延伸、扭曲、或各微流体层没有共同重合。 [0164] In the preferred embodiment, when the microfluidic device X, y coordinates of the image and the ideal plot of time, the system followed by comparing the X, y coordinates of the figure the reference mark position and the ideal image storage graph X , y coordinates of the reference marks in FIG position to determine whether there microfluidic device extends, distorted, or no layer microfluidic respective common overlap. 如果在实际基准位置和成像基准位置之间有差异,则执行矩阵变换(最好是仿射变换),W便将测定单元的成像形状变换成理想测定单元形状的虚拟形状。 If there are differences between the actual reference position and the imaging reference position, performing matrix transform (preferably an affine transformation), forming the shape of W put the measurement unit is converted into an ideal shape measurement unit virtual shape. 通过使用矩阵变换(该矩阵变换是从测得的实际基准位置和所存储的理想基准位置之间的差异中计算出来的)将实际图像转换成已知且固定的理想图像,便可W使图像减影和其它图像分析成为可能。 By using the matrix transformation (the transformation matrix is calculated from the difference between the actual and reference position measured over the reference position between the stored out of) the actual image is converted into a known and fixed over the image, you can make the image W subtraction and other image analysis possible.

[01化]通过矩阵分析计算坐标图之间的差异,矩阵变换可W演化成将实际图像重新构造成理想的图像W便作进一步的图像处理。 [Of 01] matrix by the difference between the analysis and calculation graph, matrix transformation W can evolve into actual images reconstructed as an ideal image W will further image processing. 通过使成像的微流体器件与标准形状一致,图像减影和掩模便可能使测定单元反应室的可视区域最大化。 By microfluidic device and standard imaging consistent shape, image subtraction and masking can make to maximize the viewable area measurement unit of the reaction chamber. 此外,如果在一系列基于时间的图像中时间零处的反应室内存在缺陷或碎片,则在接下来的图像中可W遮掉运种缺陷或碎片W避免施加自动分析时的伪信号。 In addition, if there are defects or debris in a series of image-based time in the reaction chamber at time zero, then the next image can cover off W transported species defects or debris W applied to avoid spurious signals when the automatic analyzer. 除了遮掉反应室中含缺陷或碎片的区域,还可W从接下来的图像中减去反应室的壁,同样是为了在接下来的分析中不引起伪读数。 In addition to cover off region of the reaction chamber with defects or debris, but also the reaction chamber wall W subtracted from the next image, the same is not to cause pseudo readings in the following analysis. 基于控制层中发现的物体(比如控制线自身)的位置,也可W计算各层之间(比如控制层和通道层之间)的差异。 Control layer based on found objects (such as self-control line) position, W may also calculate the difference between the layers (such as the control layer and the channel layer) is. 在另一个示例中,运种校正是基于控制层基准自身而确定的。 In another example, the control layer is transported species correction determined based on the reference itself. 对于某些实施例,运种额外的变换是重要的,因为控制层将蛋白质反应室与其它控制线隔离开。 For some embodiments, additional types of transport transformation is important, because the control layer protein reaction chamber and the other control lines isolated. 阳166] 图2A-2C是示出了本发明一实施例的透镜组件的简化示意图。 Yang 166] Figures 2A-2C is a simplified schematic diagram illustrating a lens assembly according to an embodiment of the present invention. 如图2A-2C所示, 每张图都提供了微流体器件205。 As shown in FIG. 2A-2C, each figure provides a microfluidic device 205. 尽管在图2A-2C中没有示出热控制器和其它系统元件,但是本领域的技术人员将会理解运两种图之间的关系。 Although not shown, the thermal controllers and other system components in FIG. 2A-2C, but those skilled in the art will understand the relationship between transport both feature. 因此,在合适的地方,标号都是取自图1A,W进一步澄清各图之间的关系。 Therefore, where appropriate, reference is taken from Fig. 1A, W to further clarify the relationship between the figures. 阳167] 图2A示出了第一透镜系统210,其中包括第一孔径211。 Male 167] Figure 2A illustrates a first lens system 210, which includes a first aperture 211. 如图所示,第一孔径定位于单独的透镜元件之间,然而,运不是本发明所要求的。 As shown, a first aperture positioned between the individual lens elements, however, operation of the present invention is not required. 在其它光学设计中,选择第一孔径的位置、大小和其它特征,W实现预定的光学设计目标。 In other optical design, selecting a first aperture position, size, and other characteristics, W to achieve a predetermined optical design goals. 图2A还示出了第二透镜系统212, 其中包括第二孔径213。 Figure 2A also shows a second lens system 212, which includes a second aperture 213. 如结合第一透镜系统210所讨论的那样,光学元件都根据预定的光学设计目标来排列。 As with the first lens system 210 as discussed, the optical elements are designed in accordance with a predetermined optical targets arranged.

[0168] 参照图IA和2A-2C,示出了和滤光器件有关的其它细节。 [0168] Referring to FIG. IA and 2A-2C, shows and other details related to the filter device. 通常,透镜系统210、212 W及滤光设备213都设置在成像通路的一部分中。 Typically, the optical lens system and equipment 210,212 W 213 are provided in a part of the imaging path. 在一些实施例中,成像通路是一种发射通路,用于将来自微流体器件的一个或多个巧光发射信号传输到检测器。 In some embodiments, the imaging path is a transmission path for a microfluidic device from the one or more light emitting signal to the coincidence detector. 如下文更全面的描述,滤光器件适用于使一个或多个巧光发射信号中的选定光谱带宽通过,并且还适用于将与一个或多个巧光发射信号相关联的一种或多种色差处理到确定的水平。 As more fully described, the filter device suitable for making one or more clever light emitting signals through the selected spectral bandwidth, and also applied to a signal associated with one or more of the light emitted or clever kinds of color processing to determine the level. 在一些实施例中,处理一种或多种色差包括减小运种色差 In some embodiments, treatment comprises reducing one or more of the color difference color difference operation

[0169] 在特定的实施例中,如图2A所示,提供了第一光谱滤光片214和零屈光度光学元件261。 [0169] In a particular embodiment, shown in Figure 2A, a first spectral filter 214 and the refractive optical element 261 is zero. 参照图1,当滤光器件213(在一些实施例中是发射滤光轮)处于第一工作位置时, 在微流体器件和检测器之间的光路中存在滤光片/零屈光度元件的组合。 When referring to FIG. 1, when the optical device 213 (in some embodiments, the emission filter wheel) is in the first operating position, the combination filter / zero refractive optical element present in the microfluidic device and the detector between . 针对大约位于光谱中屯、的波长的传输和聚焦,使图2A的光学元件最佳化。 For some of the spectrum in the village, the wavelength of the transmission and focusing of the optical element of FIG. 2A of the best. 因此,在一个实施例中,光谱滤光片214 W 570纳米波长为中屯、,该波长和巧光团fluorophore Vic (从加州化Ster市Applied Biosystems公司那儿可W买到)有关。 Thus, in one embodiment, the spectral filter 214 W 570 nm to Tuen ,, the wavelength of light and clever group fluorophore Vic (from Ster City, California-based company Applied Biosystems there can W available) related. 另外,该光谱滤光片的特征在于其30纳米的光谱带宽。 Furthermore, the spectral filter is characterized in that the spectral bandwidth of 30 nm. 在一些实施例中,如下文更全面地描述那样,还提供了适用于校正色差的滤光片/零屈光度元件,但W其它波长(通常比570纳米要短或长)为中屯、。 In some embodiments, as more fully described above, also it provides a suitable color correction filter / zero diopter element, but other wavelengths W (usually higher than 570 nm or shorter length) in Tuen.

[0170] 如结合图1中的光谱滤光片244所讨论的那样,滤光器件并不限于轮子运种形状。 [0170] connection with FIG. 1 as discussed spectral filter 244, the filter device is not limited to the transport wheel shapes. 例如,在本发明的一些实施例中,包括光谱滤光片、像差校正元件及其组合(它们落在滤光片支架中)。 For example, in some embodiments of the present invention, the spectral filter comprises, aberration correcting element, and combinations thereof (they come within the filter holder). 此外,根据本发明的实施例,还包括与像差校正元件组合在一起的电光滤波器和被驱动器置入光路中的滤光片。 Further, according to the embodiment of the invention, further comprising in combination with the aberration correcting element and the electro-optical filter together drive the filter into the light path. 阳171] 图2B示出了第一和第二透镜系统,其中发射滤光轮位于第二工作位置。 Male 171] Figure 2B shows a first and a second lens system, wherein the emission filter wheel is in the second operating position. 在图2B 所示的实施例中,光谱滤光片224 W 518纳米波长为中屯、(该波长与巧光团Fam™有关,可W从Applied Biosystems公司买到)并且其特征在于25纳米的光谱带宽。 In an embodiment, the spectral filter shown in Figure 2B 224 W 518 nanometer wavelength in the village, (the wavelength of light and clever about the group Fam ™, available from Applied Biosystems. W available) and is characterized by 25 nm spectral bandwidth. 通常,光谱滤光片224适用于透射和在"蓝色"波长处发光的巧光团有关的巧光信号,通常该巧光信号和光谱短波长部分附近的波长相关。 Typically, the spectral filter 224 and is suitable for transmission in the "blue" wavelength light emitting Qiao Qiao groups associated optical signal, usually related to the clever optical signal wavelength and spectral close to short-wavelength part.

[0172] 图2B示出了充当零屈光度双透镜的光学元件226。 [0172] Figure 2B shows a dual function as a zero diopter lens 226 of the optical element. 在一些实施例中,滤光片/零屈光度双透镜是作为复合光学元件而提供的,而在其它实施例中,滤光片和零屈光度双透镜是彼此分开的。 In some embodiments, the filter / zero diopter lens pairs are provided as a compound optical element, whereas in other embodiments, the filter and zero diopter lens pairs are separated from each other. 此外,在一些实施例中,使发射滤光轮旋转W调整滤光片/零屈光度光学元件的位置,从而从第一工作位置变到第二工作位置。 Further, in some embodiments, the emission filter wheel so that rotation of the adjustment Filter W / zero diopter position of the optical element, so that changes from a first operative position to a second working position. 图2B所示的零屈光度双透镜226被设计成用于校正由光学系统在"蓝色"波长处引入的色差。 Figure 2B double zero diopter lens 226 is designed to be shown by the optical system for correcting the "blue" wavelengths chromatic aberration introduced. 在特定的实施例中,选择零屈光度双透镜,W便校正与特定巧光团(比如Fam™)的发光相关的波长处的色差。 In a particular embodiment, the select zero double refraction lens, chromatic aberration correction W will light and clever specific groups (such as Fam ™) emitting at a wavelength associated.

[0173] 在一些实施例中,零屈光度双透镜是由多种折射率数值不同的单独的光学材料制成的。 [0173] In some embodiments, the double zero diopter lens is a combination of different refractive index values of the individual optical material. 仅作为示例,如图2B所示,平凹透镜禪合到凸平透镜。 Of example only, 2B, Zen plano-concave lens bonded to the flat convex lens. 在特定的实施例中,零屈光度双透镜是一种由多个光学元件胶合在一起的夫巧和费消色差透镜。 In a particular embodiment, the double zero diopter lens is a cemented together by a plurality of optical elements and clever husband fees achromatic lens. 在其它实施例中,使用对本领域技术人员而言很明显的可替换的设计。 In other embodiments, the use of skilled in the art will be apparent to an alternative design. 在一些实施例中,使用本说明书中所示的嵌入式双透镜,来减小蓝色和红色滤光波带中的轴向色差。 In some embodiments, the use of embedded dual lens shown in the present specification, the filter to reduce the blue and red band in the axial chromatic aberration.

[0174] 图2C示出了第一和第二透镜系统,其中发射滤光轮处于第=工作位置。 [0174] Figure 2C shows a first and a second lens system, wherein the emission filter wheel is in the first working position =. 在图2C 所示的实施例中,光谱滤光片228 W 645纳米波长为中屯、(该波长和巧光团Rox™有关,运种巧光团可W从Applied Biosciences公司买到)并且特征在于75纳米的光谱带宽。 In the embodiment shown in FIG. 2C, spectral filter 228 W 645 nanometer wavelength in the village, (the wavelength of light and clever group Rox ™ related transport kinds of clever light W group may buy the company from Applied Biosciences) and features that the spectral bandwidth of 75 nm. 通常,光谱滤光片228适用于透射与在"红色"波长处发光的巧光团有关的巧光信号,该巧光信号通常与光谱的长波长部分附近的波长有关。 Typically, the spectral filter 228 is suitable for optical signal transmission and clever in the "red" light at the wavelength of the light emitting clever about the group, which is usually clever and long-wavelength optical signal wavelength spectrum portion near about.

[01巧]如图2B所示,图2C示出了充当零屈光度双透镜的光学元件230。 [Qiao 01] shown in Figure 2B, Figure 2C shows a dual function as a zero diopter lens 230 of the optical element. 在一些实施例中,滤光片/零屈光度双透镜是作为复合光学元件来提供的,而在其它实施例中,滤光片和零屈光度双透镜彼此分开。 In some embodiments, the filter / zero diopter lens pairs are provided as a compound optical element, whereas in other embodiments, the filter and zero diopter lens pairs separated from each other. 此外,在一些实施例中,使发射滤光轮旋转,W便调整滤光片/ 零屈光度光学元件的位置,从而从第一或第二工作位置变为第=工作位置。 Further, in some embodiments, the emitter rotating filter wheel, W are adjusted filter / zero diopter position of the optical element, whereby the first or the second working position to the working position = first. 图2C所示的零屈光度双透镜230被设计成校正由光学系统在"红色"波长处所引入的色差。 Figure 2C double zero diopter lens 230 is shown by the optical system is designed to be corrected in the "red" wavelength chromatic aberration introduced premises. 在特定的实施例中,选择零屈光度双透镜,W便校正与特定的巧光团(例如Rox™)发光有关的波长处的色差。 In a particular embodiment, the select zero double refraction lens, W will be corrected with a particular clever light group (for example Rox ™) color of light emission at a wavelength of about. 比较图2B和2C,图2C所示的零屈光度双透镜包括禪合到凹平透镜的平凸透镜。 Comparing Figures 2B and 2C, Fig. 2C double zero diopter lens shown includes Zen fit into the recess flat lens plano-convex lens.

[0176] 如结合图2A、2B和2C所讨论的那样,与各个滤光片有关的校正光学元件216、226 和230被设计成,它们帮助校正在上述各个滤光片所透射的波长处的色差。 [0176] The combination of Figures 2A, 2B and 2C as discussed, the respective correction optical element 216, 226 and associated filter 230 are designed so that they help in correcting each of said filter wavelength of the transmitted color. 运种设计允许在不同的滤光片(透射不同的波长区域)上使透镜系统的光斑大小、模糊和其它光学特性更均匀和更一致。 Yun design allows different types of filters (transmission in different wavelength regions) so that the spot size of the lens system, fuzzy and other optical characteristics more uniform and more consistent. 运些益处可用于许多应用,例如,微流体器件中的反应室紧密填充。 These benefits can be used to transport a number of applications, e.g., microfluidic device tightly packed in the reaction chamber. 另夕F,针对来自不同反应室的光信号之间的可允许串扰而定的设计目标通常对图像平面上可允许的最大光斑或模糊的尺寸给出限制。 Another evening F, for allowable crosstalk given design objective optical signals from different reaction chambers between the usual spot on the image plane of the maximum allowable limit or obscure dimensions given. 像差校正光学元件减小一个波长极限(例如,蓝色波长区域中的滤光透射)处的模糊或光斑大小,并且还减小不同波长区域处的模糊或光斑大小(例如,对于其通带位于红色波长区域中的滤光片)。 Aberration correction optical element is reduced a wavelength limit (for example, the blue wavelength region of the optical transmission) or blur spot size at, and also to reduce blur or spot size at a different wavelength region (for example, for passband of the red filters in the wavelength region). 该益处可用于下列情形:整个测量是一种比例度量,运取决于在红色和蓝色波长区域中检测到的单独信号,由此提高了整个试验的灵敏度。 The benefit can be used in the following situations: The entire measurement is a measure of the ratio, depending on the operation of individual signals detected in the red and blue wavelength regions, thereby increasing the sensitivity of the entire trial. 上文提供的讨论设及特定的实施例,但是本领域的技术人员应该理解, 有许多校正零屈光度光学元件的变体,它们W相似的方式和具有不同波长通带的滤光片结合起来被置于光学系统中,运些都被包括在本发明的范围中。 And disposed above discussion provides specific embodiments, those skilled in the art will appreciate, there are many variations of the zero diopter correction optical element, in a similar manner and their W filters having different pass wavelength bands are combined placed in the optical system, the transport of these are included in the scope of the present invention.

[0177] 图3是本发明一实施例的微流体器件中存在的许多反应室中发生的反应所产生的W第一波长为中屯、的巧光发射的照片。 [0177] FIG. 3 is a first wavelength W react many reaction chamber microfluidic device of one embodiment of the present invention is present in the produced in the village, the light-emitting clever photos. 如图3所示,许多反应室产生第一波长"波长1" 的发光。 As shown in Figure 3, many of the reaction chamber to produce a first wavelength "wavelength 1" luminescence. 在图3所示的本发明的实施例中,10x16的反应室阵列被成像。 In the embodiment shown in FIG. 3 of the present invention, the reaction chamber 10x16 array is imaged. 在一些实施例中, 第一波长与上述巧光团相关联。 In some embodiments, the first wavelength and said light group associated with skillfully. 对于本领域的技术人员而言,很明显,第一波长的发光强度是反应室中发生的化学过程的函数。 For those skilled in the art, it is quite clear that the emission intensity of the first wavelength is a function of the chemical processes occurring in the reaction chamber. 如图所示,用根据本发明一实施例提供的光学成像系统给10x16反应室的二维阵列成像。 As shown, to a two-dimensional array of imaging 10x16 reaction chamber optical imaging system provided in accordance with an embodiment of the invention. 如上所述,在本发明的一些实施例中,反应室处于流体隔离。 As described above, in some embodiments of the present invention, the reaction chamber is fluidly isolated. 此外,根据可替代的实施例,反应室的特征在于,体积在纳升范围,和/或反应室密度的量级为每平方厘米有几百个反应室。 Further, according to an alternative embodiment, the reaction chamber is characterized in that the volume in the nanoliter range, and / or density of the reaction chamber is of the order a few hundred square centimeters per reaction chamber.

[0178] 图4是本发明一实施例的微流体器件中存在的许多反应室中发生的反应所产生的W第二波长为中屯、的巧光发射的照片。 [0178] FIG. 4 is a second embodiment of the present invention W wavelength reacts many microfluidic device according to the reaction chamber in the presence of the village is generated, the light-emitting clever photos. 如图4所示,许多反应室产生第二波长"波长2" 处的发光比较图3和4, 一些反应室产生很少的光或不产生光,而其它反应室产生第一波长的光、第二波长的光、或第一和第二波长的光。 As shown in Figure 4, many of the reaction chamber 4 and generates a second wavelength, some of the reaction chamber to generate a first wavelength of light or no light, while the other chamber to produce very small "wavelength 2" at the light emitting Comparing Figure 3, a second wavelength, or the first and second wavelengths. 当然,巧光活性的收集和分析可W对反应室中发生的化学反应的本质作出理解。 Of course, the collection and analysis of clever photoactive W can be the essence of chemical reactions take place in the reaction chamber to understand. 阳179] 图5-7是用本发明一实施例产生的选定波长的光点图。 Yang 179] Figures 5-7 with the present invention, a selected wavelength of the light spot generated view of implementation. 参照运些图的说明,图中所示的波长范围通常分成=个波带:绿色波长,蓝色波长和红色波长。 Referring to FIG shipped some explanation, the wavelength range shown in the figure is usually divided into = a band: a green wavelength, blue wavelength and the red wavelength. 图5示出了525、550 和575nm的波长,它们通常与光谱的绿色区域相关联。 Figure 5 shows a 525,550 and 575nm wavelengths, they usually green region of the spectrum is associated. 图6示出了486、501和516nm的波长,它们通常与光谱的蓝色区域相关联。 Figure 6 shows a 486,501 and 516nm wavelengths, they usually associate blue region of the spectrum. 图7示出了616、636和656nm的波长,它们通常与光谱的红色区域相关联。 Figure 7 shows a 616,636 and 656nm wavelengths are typically red region of the spectrum is associated. 图5是针对图2A所示的透镜系统而计算的,图6是针对图2B所示的透镜系统而计算的,而图7是针对图2C所示的透镜系统而计算的。 Figure 5 is directed to the lens system shown in FIG. 2A and calculated for the lens system 6 is shown in Figure 2B calculated, and Figure 7 for the lens system shown in Figure 2C calculated. 阳180]图8是用本发明一实施例产生的照度图,该图示出了作为位置的函数的相对均匀性。 Yang 180] FIG. 8 is an embodiment of the present invention produces illumination diagram, which shows the relative position as a function of the uniformity. 在图8中,针对图2A所示的光学系统,绘出了作为Y段(毫米)的函数的相对照度。 In Figure 8, for the optical system shown in FIG. 2A, depicted as segment Y (mm) as a function of the degree of contrast. 在0. 550 ym的波长处,超过距离21. 4mm的相对照度均匀性大于90%。 0. 550 ym in wavelength, than the distance 21. 4mm illuminance uniformity of greater than 90%. 阳181] 图9-11是本发明若干实施例的正方块块内能量图。 Yang 181] 9-11 are within the present invention, a plurality of square tranches energy diagram of an embodiment. 在一些光学系统中,光学系统性能的测量是正方块块内能量百分比,运是指定中屯、区域中的总能量的百分比。 In some optical systems, the measurement of the optical performance of the system is the percentage of energy within the square tranches, transport is specified in Tuen, the percentage area of the total energy. 参照图9, 通过使用图2A所示的透镜系统,所包括的能量的百分率被绘制成各位置的形屯、的半宽(W 微米为单位)的函数。 Referring to FIG. 9, the percentage of energy by using the lens system shown in Figure 2A, including the shape Tun graphed each position, the half-width (W microns) function. 作为一个示例,对于离中屯、14. 8mm的位置,约50%的能量被包括在离形屯、约7. 5 ym处,而对于离中屯、21. 4mm的位置,约90%的能量被包括在离形屯、相同距离处。 As an example, for from the village, 14. 8mm location, about 50% of the energy is included in shape from the village, at about 7. 5 ym, and for off Tuen, 21. 4mm location, about 90% energy is included in shape from the village, at the same distance. 在图9-11中,包括衍射,通常用快速傅立叶变换(FFT)算法来执行上述计算。 In Figures 9-11, including diffraction, usually Fast Fourier Transform (FFT) algorithm to perform the above calculations. 图10 和11分别是图2B和2C所示透镜系统的正方块块内能量图。 Figures 10 and 11 respectively in Fig. 2B and 2C, the lens system within a square tranches Energy. 在运些图中,如图9所示,包括衍射。 In operation these figures, shown in Figure 9, including diffraction. 阳182]图12是示出了本发明一实施例所提供的光学系统的场曲和崎变的图。 Yang 182] FIG. 12 is a diagram showing the embodiment of the invention the curvature of field provided by the optical system and of distortion according to one example. 针对波长0. 525 ym、0. 550 ym和0. 575 ym示出了场曲,它们都标记在图中。 For wavelength 0. 525 ym, 0. 550 ym and 0. 575 ym shows a field curvature, they are marked in the figure. 对于不同的所示波长,崎变不相同,其中各波长之间可忽略的差异在0.05%的水平。 For different wavelengths, the change is not the same as Kawasaki, wherein between each wavelength negligible differences in the level of 0.05%. 阳18引图13是本发明一实施例的系统所产生的"双波长-焦点"的图。 Male 18 lead 13 is a system embodiment of the invention is produced by the "dual wavelength - focus" FIG. 在该图中,色焦点移动是波长(W微米为单位)的函数。 In the figure, the color is the focus movement wavelength (W microns) function. 图13所示的波长范围覆盖了从480纳米到650纳米的波长范围。 The wavelength range shown in FIG. 13 covers from 480 nm to 650 nm wavelength range. 如图所示,最大焦点移动范围是214. 3702 ym,而衍射限制范围是4. 243 ym。 As shown, the maximum range of movement of the focus is 214. 3702 ym, and diffraction limited range of 4. 243 ym. 参照图13,超过214 ym长的颜色焦点跨越所示范围。 Referring to FIG. 13, more than the 214 ym focal length of color across the range indicated. 因此,系统性能的分析通常包括要考虑+IOOym的散焦处的光斑大小。 Therefore, the analysis of system performance typically include consideration at + IOOym defocused spot size. 通常,散焦光斑的大小将超过焦点中的光斑的大小。 Typically, defocused spot size will exceed the size of the focus spot. 根据本发明各实施例,通过使用上述嵌入式双透镜,来校正轴向色差。 According to various embodiments of the present invention, by using the embedded double lens, the axial chromatic aberration is corrected. 在可选的实施例中,使用特殊的玻璃类型来实现复消色差性能(通常在比其它玻璃类型要贵的情况下获得)。 In an alternative embodiment, the use of special types of glass to achieve apochromatic performance (usually at expensive than other types of glass obtained in a case). [0184] 图14-16是用本发明一实施例产生的针对选定波长的光点图。 [0184] FIG. 14-16 is the wavelength of the light spot diagram for the selected embodiment of the present invention to produce an embodiment. 如图14的说明所示,当上述系统在最佳焦点位置时,提供了光谱绿色区域中若干个波长巧25、550和575nm) 的光点图。 Description As shown in Figure 14, when said system is in the best focus position, provides a green spectral region several wavelengths clever 25,550 and 575nm) light spots in FIG. 为了产生图14-16所示的光点图,使用了图2A所示的透镜系统。 In order to produce the light spot shown in Figure 14-16, the use of the lens system shown in Figure 2A. 图15和16分别是在+100 ym的散焦和-100 ym的散焦处计算出的光点图。 Figures 15 and 16 respectively at +100 ym defocus and -100 ym defocus calculated at the spot diagram. 在图15和16中,考虑光谱绿色区域中相同的波长。 In Fig. 15 and 16, in consideration of the green spectral region of the same wavelength. 阳185] 图17-19是用本发明一实施例产生的针对选定波长的光点图。 Yang 185] 17-19 is an embodiment of the present invention is produced for the selected wavelengths of light dot plot. 如图17的说明所示,当系统位于最佳焦点位置时,提供了光谱蓝色区域中若干个波长(486、501和516nm)的光点图。 Description As shown in FIG. 17, when the system is located in the best focus position, provides a spectrum of several wavelengths in the blue region (486,501 and 516nm) light spots in FIG. 为了产生图17-19所示的光点图,使用了图2B所示的透镜系统。 In order to produce the light spot shown in Figure 17-19, the use of the lens system shown in Figure 2B. 图18和19分别是在+100 ym的散焦和-100 ym的散焦处计算的光点图。 Figures 18 and 19 are in the spot diagram +100 ym defocus and -100 ym defocus calculated at. 在图18和19中,考虑光谱的蓝色区域中相同的波长。 In Fig. 18 and 19, taking the blue region of the spectrum the same wavelength. 如图19所示,场4包括501皿处的光线,延伸到IOOym盒子W外。 19, field 4 includes light dish at 501, extending to IOOym W outside the box.

[0186] 图20-22是用本发明一实施例产生的针对选定波长的光点图。 [0186] FIG. 20-22 is the wavelength of the light spot diagram for the selected embodiment of the present invention to produce an embodiment. 如图20的说明所示,在系统位于最佳焦点位置时,提供了光谱的红色区域中若干个波长化16、636和656nm) 的光点图。 Description shown in FIG. 20, when the system is located in the best focus position, it provides a spectrum of several wavelengths in the red region of 16,636 and 656nm) light spots in FIG. 为了产生图20-22所示的光点图,使用了图2C所示的透镜系统。 In order to generate a light spot shown in Fig. 20-22, the use of a lens system is shown in Figure 2C. 图21和22分别是在+IOOym散焦和-IOOym散焦处计算出的。 Figures 21 and 22 respectively at + IOOym -IOOym defocus and defocus calculated at. 在图21和22中,考虑光谱的红色区域中相同的波长。 In Fig. 21 and 22, taking the red region of the spectrum in the same wavelength.

[0187] 在本发明的可选实施例中,提供了另一个1 : 1光学中继成像系统,其中包括对图2A-2C所示光学元件的修改。 [0187] In an alternative embodiment of the present invention, there is provided another 1: 1 relay optical imaging system, including modification of the optical element shown in Figures 2A-2C. 尽管保留了一般的光学系统,但是特定元件(包括滤光片/零屈光度双透镜的组合)的特性被修改了。 Despite the reservations of the general optical system, a particular element (including a combination filter / double zero diopter lens) features were modified. 在运种可选实施例中,提供了大于35mm(例如,46. 12mm)的工作距离。 In an alternative embodiment kind of operation, providing greater than 35mm (e.g., 46. 12mm) working distance. 此外,提供了平均为11.28 ym的RMS光斑直径,其中最大值是14. 73 ym,光学成像系统的总长是542. 2mm,其中最大滤光片AOI为12. 59度,且滤光片处的最大光束直径为76mm。 In addition, there is provided an average of 11.28 ym of RMS spot diameter, where the maximum is 14. 73 ym, the total length of the optical imaging system is 542. 2mm, the largest filter AOI is 12.59 degrees, and filters at the the maximum beam diameter of 76mm.

[0188] 图23-25是用本发明一可选实施例产生的针对选定波长的光点图。 [0188] FIG. 23-25 of the present invention is a wavelength-selective optical dot pattern generated for the selected example embodiments. 参照运些图的说明,图中所示波长范围通常分成=个波带:绿色波长,蓝色波长,和红色波长。 Referring to FIG shipped some explanation, as shown in FIG wavelength range = typically divided into a band: a green wavelength, blue wavelength, and red wavelength. 图23示出了波长525、550和575nm,它们通常和光谱的绿色区域相关。 Figure 23 shows a wavelength 525,550 and 575nm, and they are usually related to the green region of the spectrum. 图24示出了波长486、501和516nm,它们通常和光谱的蓝色区域相关。 Figure 24 shows a wavelength 486,501 and 516nm, and they are usually related to the blue region of the spectrum. 图25示出了波长621、636和651nm,它们通常和光谱的红色区域相关。 Figure 25 shows a wavelength 621,636 and 651nm, and they are usually related to the red region of the spectrum. 图23是基于图2A针对透镜系统而计算的,图24是基于图2B针对透镜系统而计算的,图25是基于图2C针对透镜系统而计算的。 FIG. 23 is based on FIG. 2A and the lens system for the calculation, based on FIG. 24 is a diagram for lens systems 2B calculated, FIG. 25 is a diagram for 2C calculated based on the lens system.

[0189] 图26-28是根据本发明的若干实施例的正方块块内能量图。 [0189] FIG. 26-28 square tranches within an energy diagram according to several embodiments of the present invention. 参照图26,通过使用图2A所示的透镜系统,针对各个位置,绘出了所包括的能量的百分率,作为形屯、半宽(W微米为单位)的函数。 Referring to FIG. 26, by using the lens system shown in Figure 2A, for each position, the percentage of energy drawn included, as Tun-shaped, half-width (W microns) function. 作为一个示例,对于离中屯、14. 8mm的位置,约80%的能量被包括在离形屯、约7. 5 ym处,而对于离中屯、21. 4mm的位置,约90 %的能量被包括在离形屯、相同距离处。 As an example, for from the village, 14. 8mm location, about 80% of the energy is included in shape from the village, at about 7. 5 ym, and for off Tuen, 21. 4mm location, about 90% energy is included in shape from the village, at the same distance. 在图26-28中,包括衍射,通常用快速傅里叶变换(FFT)算法来执行计算过程。 In Figure 26-28, including diffraction, usually Fast Fourier Transform (FFT) algorithm to perform the calculation process. 图27 和28分别是图2B和2C所示透镜系统的正方块块内能量图。 27 and 28 are within the square tranches FIG. 2B and 2C lens system energy diagram. 在运些图中,像在图26中那样,包括衍射。 In transport these figures, like that, including diffraction in Fig. 26.

[0190] 还应该理解,上文所描述的示例和实施例只用于解释说明,并且各种修改和变化对于本领域的技术人员而言都是很明显的并且应该被包括在本申请的精神和权限W及所附权利要求书的范围之内。 [0190] It should also be understood that the examples described above, and explanation of examples only, and various modifications and variations to persons skilled in the art and are apparent should be included in the spirit of this application and authority within the W and the scope of the appended claims.

Patent Citations
Cited PatentFiling datePublication dateApplicantTitle
CN1460855A *13 Jun 200310 Dec 2003中国科学院生物物理研究所Microfluid biological sensor chip device and its application
CN1534312A *30 Mar 20046 Oct 2004朗迅科技公司Method and device for controlling liquid motion on surface of ultrafine structure or microstructure
Legal Events
DateCodeEventDescription
4 Aug 2010C06Publication
22 Sep 2010C10Entry into substantive examination
13 Apr 2016C14Grant of patent or utility model