CN100414288C - Miniature millimeter laser induced fluorescent detector for biological chip - Google Patents
Miniature millimeter laser induced fluorescent detector for biological chip Download PDFInfo
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- CN100414288C CN100414288C CNB200510087115XA CN200510087115A CN100414288C CN 100414288 C CN100414288 C CN 100414288C CN B200510087115X A CNB200510087115X A CN B200510087115XA CN 200510087115 A CN200510087115 A CN 200510087115A CN 100414288 C CN100414288 C CN 100414288C
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Abstract
The present invention relates to a millimetre-level miniature laser-induced fluorescence detection instrument for biological chips, which belongs to biology and medicine detection instruments. The millimetre-level miniature laser-induced fluorescence detection instrument comprises a light source, a light-filtering system and a photoelectric detection system and is characterized in that the light source is a semiconductor laser device (1); the peak wavelength of the semiconductor laser device (1) is from 470 to 495 nm, and the spectrum is stopped in the depth of 500 nm; the light-filtering system is an interference light-filtering film (2); the stop wavelength of the interference light-filtering film (2) is 500 nm, and the peak wavelength is 520 nm; the photoelectric detection system is a photoelectric detection component (3) of a miniature semiconductor; the semiconductor laser device, the interference light-filtering film and the photoelectric detection component are integrated; the total volume of the fluorescence detection instrument is in the level of millimetre, and the length, the width and the height of the fluorescence detection instrument are in the range of 1mm to 10mm; the photoelectric detection component can be positioned right below the semiconductor laser device, can enclose the semiconductor laser device and can be in parallel to the semiconductor laser device. The present invention simplifies the structure of the device of the detection technology, the device is convenient to carry, and the brought errors are eliminated, and thereby, the detection result is close to the true value.
Description
Technical field
The present invention mainly is aimed at the excitation wavelength peak value at 470～495nm, and emission wavelength belongs to biology and medicine detector device in the detection of the fluorescent material of 510～530nm.
Background technology
Biochip is a new and high technology that develops rapidly in life science in recent years, because biochip can be analyzed a large amount of biomolecule at short notice, make people obtain biological information in the sample rapidly and accurately, efficient is hundreds and thousands of times of traditional detection means, thereby presents the development peak in a short time.
Biochip technology has 4 bare bones: chip preparation, specimen preparation, biomolecule hybridization reaction and input.Input is the important component part of biochip technology, mainly comprises hybridization signal generation, signal collection and transmission and signal Processing and discerns three parts.In to the biochip signal detecting method, the fluorescence detection good reproducibility, selectivity is strong, adopts one of maximum method at present.
When some material is shone by the light (exciting light) of certain wavelength, in the extremely short time, can launch color immediately and intensity has nothing in common with each other and than the longer light of exciting light (also claiming absorbing light) wavelength, and disappear immediately along with the disappearance of exciting light, this kind light is fluorescence.
The ultimate principle of fluorescence detection is: the sample that will detect is with fluorescein-labelled, and fully hybridize with the known (probe) on the chip, use the image display result behind the wash-out, detect the expressed relevant biological information of testing sample by Computer Processing then.The purpose of mark is that make can detected trace labelling on the sample band to be measured.
Fluorescent material (fluorescein) produces the highest release strength at the fluorescence that is subjected to discharging after exciting light excites certain intensity under a certain wavelength, and the absorption value that excites is separately arranged.The absorption peak 480nm of a kind of fluorescent material FluorX commonly used and emission peak ripple 520nm are behind the testing sample and the hybridization of the probe array on the biochip of mark, fluorescently-labeled sample is combined on the ad-hoc location of chip, hybrid molecule is not removed, need this moment the chip measurement result to be transformed into and can to detect with the fluorescence signal of realizing biochip for the view data of analyzing and processing with pick-up unit.
Mainly be divided into two big classes based on the real-time detection technique of fluorescently-labeled biochip signal at present.
One, the laser scanning fluorescent microscope detects, laser confocal scanning microscope detection etc.Its shortcoming is: scanning accuracy mainly is subjected to the influence of mechanical precision, repeatable accuracy and the environmental baseline of X, Y mobile platform.
Two, use the signal detection technique of the weak light detection devices such as CCD of cooling.Because the excitation light irradiation light field is the entire chip zone, and the Gaussian distribution of laser beam intensity, can make light field light distribution inequality, and the fluorescence labeling signal intensity with excite light intensity linear, acquired signal can accurately linear response.CCD detects and to be based on that Flame Image Process finishes, and therefore is easy to bring distorted signals.
The structure of the device of these detection techniques of while is more complicated all, is not easy to carry.
Summary of the invention
The purpose of this invention is to provide a kind of millimeter level laser induced fluorescence detector that excitation wavelength is detected at the fluorescent material of 510～530nm at 470～495nm, emission wavelength.This millimeter level standalone probe unit both can avoid the detection system scanning accuracy mainly to be subjected to the influence of mechanical precision, repeatable accuracy and the environmental baseline of X, Y mobile platform, can guarantee the shimming irradiation of exciting light again, the more important thing is the fluorescence signal intensity that can improve the fluorescence biosensor chip mark, precision, the repeatable accuracy of scanning, the structure of simplification checkout equipment.
The millimeter level miniature laser induced fluorescence detector that biochip of the present invention uses, include light source, filter system and photodetector system, it is characterized in that: light source is the semiconductor laser 1 that peak wavelength ends in the 500nm place degree of depth at 470～495nm, its spectrum, filter system is that cutoff wavelength is the interference light filtering film 2 of 500nm peak wavelength at 520nm, and photoelectric detection part is a micro semiconductor photoelectric detector 3; Semiconductor laser 1, interference light filtering film 2, photoelectric detector 3 are integrated in one; The fluorescence detector cumulative volume is in the millimeter rank, and its length is in 1mm～10mm scope.This fluorescence detector can infinitely be pressed close to detected object, or enters and carry out fluoroscopic examination in the measurand, so fluorescence is very short from being energized into the light path that incides on the photoelectric detector, distance at tens microns to ten millimeter.Light source adopts semiconductor laser device, and collimation is better, and spectral bandwidth is very narrow, can keep away filtering system.
Interference light filtering film 2 is positioned at photoelectric detector 3 tops and the photoelectric detector 3 common semiconductor lasers 1 that surround among Fig. 3, and the semiconductor laser 1 that is positioned at central authorities is circle or rectangle, and photoelectric detector 3 is circular or polygon.
The electrode of photoelectric detector 3 adopts the indium tin oxide transparent film, increases effective light-receiving area.
Interference light filtering film 2 directly is plated in the top of sensitive detection parts, perhaps is plated in earlier on other transparent substrate, and transparent substrate is attached to above the photoelectric detector 3 again.
Interference light filtering film 2 thickness are used for filtering fluorescence excitation wave band veiling glare in addition at 1～10 μ m.
This fluorescence detection device can be formed array and form the quick scanning system of light addressing.
Principle of work of the present invention is: the semiconductor laser in the miniature laser induced fluorescence detector send and by the peak value of shaping in the light beam of 470～495nm (or not shaping, but infinitely near measurand), guarantee simultaneously to end in the 500nm place degree of depth.Fluoresceins such as FluorX send the fluorescence of peak value at 520nm under laser excitation, fluorescence is received by photoelectric detector by filter plate and realizes opto-electronic conversion, the output corresponding electric signal.
The length of this device volume can be microminiaturized, reaches in 1mm～10mm scope; A plurality of miniature luminoscopes can be formed array and use functions such as realizing fluoroscopic examination multiple spot Real-time and Dynamic, the quick scanning of light addressing.In scanning detects, realize that light source, measurand and photodetector system do not have relative motion, eliminated error and the distortion that displacement brings and made testing result more near true value.Replace existing macroscopical instrument, as the PCR quantitative and qualitative detection system of general use plastics reaction tube.
Description of drawings
Fig. 1 device of the present invention (photoelectric detector is positioned under the semiconductor laser) synoptic diagram;
Fig. 2 device of the present invention (photoelectric detector and semiconductor laser are arranged side by side) synoptic diagram;
Fig. 3 device of the present invention (semiconductor laser is surrounded by photoelectric detector) synoptic diagram;
Fig. 4 the preferred embodiments of the present invention;
Fig. 5 the preferred embodiments of the present invention array combination is used for the PCR Real-time and Dynamic Detection;
Among Fig. 1-5,1, semiconductor laser device 2, interference light filtering film 3, photoelectric detector 4, fluorescence detector carrier; 5, sheet 7, microchannel 8 under biochip last slice 6, the biochip, contain fluorescein FluorX solution 9, emission light 10, fluorescence
Embodiment
With reference to accompanying drawing 4-5, will be described in detail the preferred embodiments of the present invention.
With reference to accompanying drawing 4: two 5 up and down of biochips, 6 adopt the PMMA pmma materials to make, and every organic glass thickness is 1mm.Adopt the excimer laser process technology to process the microchannel in the above.Adopt the thermocompression bonding mode with two 5,6 permanent closure about the biochip.Light source is blue-light semiconductor Laser Devices 1, peak wavelength is at 473nm, its emission light 9 shines and contains fluorescein FluorX solution 8 in the biochip microchannel 7, the fluorescence 10 that is excited is by received by photoelectric detector 3 after the filtration of interference light filtering film 2, be converted to corresponding electric signal through photoelectric detector by light signal, discern fluorescence signal intensity with this.
With reference to accompanying drawing 5: a plurality of miniature luminoscopes can be formed array and use and realize PCR fluoroscopic examination multiple spot Real-time and Dynamic.Real-time quantitative RT-PCR (Real-time reverse transcriptionquantitative polymerase chain reaction, Real-time RT-PCR), so-called real-time quantitative PCR is meant during the amplification of PCR index measures the amount of specificity product immediately by the power of continuous detecting fluorescence signal, and infers the original bulk of genes of interest in view of the above.Each circulation of passage on the pcr chip is through sex change, annealing and extend three warm areas.DNA chain of every amplification just has a fluorescence molecule to form, and can realize that the accumulation of fluorescence signal and PCR product form fully synchronously.The optical characteristics of each fluorescence signal can be by miniature luminoscope identification.
Each miniature luminoscope corresponds to the zone that will survey in each circulation of PCR respectively, be full of fluorescein FluorX solution 8 in the microchannel 7 of biochip, the emission light 9 of semiconductor laser device 1 shines the luciferin solution 8 in the biochip microchannel 7, the fluorescence 10 that is excited is by received by photoelectric detector 3 after the filtration of interference light filtering film 2, be converted to corresponding electric signal through photoelectric detector by light signal, can infer the feature of each circulation products of PCR according to the different output valves of electric signal.
Claims (9)
1. the millimeter level miniature laser induced fluorescence detector that uses of biochip, include light source, filter system and photodetector system, it is characterized in that: light source is the semiconductor laser (1) that peak wavelength ends in the 500nm place degree of depth at 470～495nm, its spectrum, filter system is that cutoff wavelength is the interference light filtering film (2) of 500nm peak wavelength at 520nm, and photoelectric detection part is micro semiconductor photoelectric detector (3); Semiconductor laser (1), interference light filtering film (2), photoelectric detector (3) are integrated in one; The fluorescence detector cumulative volume is in the millimeter rank, and its length is in 1mm～10mm scope.
2. the millimeter level miniature laser induced fluorescence detector that biochip according to claim 1 uses, it is characterized in that: photoelectric detector (3) be positioned at semiconductor laser (1) under, between photoelectric detector (3) and the semiconductor laser 1 interference light filtering film (2), semiconductor laser (1) and photoelectric detector (3) are positioned at the same side of detected object, semiconductor laser device is positioned at the centre of sensitive detection parts, and the photosensitive area area of photoelectric detector (3) is 2～50 times of semiconductor laser (1) cross-sectional area.
3. the millimeter level miniature laser induced fluorescence detector that biochip according to claim 1 uses is characterized in that: semiconductor laser (1) be arranged on interference light filtering film (2) above the photoelectric detector and photoelectric detector (3) side by side towards detected object.
4. the millimeter level miniature laser induced fluorescence detector that biochip according to claim 1 uses, it is characterized in that: interference light filtering film (2) is positioned at photoelectric detector (3) top, surround semiconductor laser (1) jointly with photoelectric detector (3), the semiconductor laser (1) that is positioned at central authorities is circle or rectangle, and photoelectric detector (3) is circle or polygon.
5. according to the millimeter level miniature laser induced fluorescence detector of claim 1 or 2 or 3 or 4 described biochips uses, it is characterized in that: the electrode of photoelectric detector (3) adopts the indium tin oxide transparent film, increases effective light-receiving area.
6. the millimeter level miniature laser induced fluorescence detector that uses according to claim 1 or 2 or 3 or 4 described biochips, it is characterized in that: interference light filtering film (2) directly is plated in the top of sensitive detection parts, perhaps be plated in earlier on other transparent substrate, transparent substrate is attached to above the photoelectric detector (3) again.
7. according to the millimeter level miniature laser induced fluorescence detector of claim 1 or 2 or 3 or 4 described biochips uses, it is characterized in that: interference light filtering film (2) thickness is at 1～10 μ m.
8. according to the millimeter level miniature laser induced fluorescence detector of claim 1 or 2 or 3 or 4 described biochips uses, it is characterized in that: a described millimeter level miniature laser induced fluorescence detector can be formed array and form the quick scanning system of light addressing.
9. according to the millimeter level miniature laser induced fluorescence detector of claim 1 or 2 or 3 or 4 described biochips uses, it is characterized in that: adopt fluorescence detector carrier (4) to be used for fixing semiconductor laser (1) and photoelectric detector (3).
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| Application Number | Priority Date | Filing Date | Title |
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| CNB200510087115XA CN100414288C (en) | 2005-07-27 | 2005-07-27 | Miniature millimeter laser induced fluorescent detector for biological chip |
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| CNB200510087115XA CN100414288C (en) | 2005-07-27 | 2005-07-27 | Miniature millimeter laser induced fluorescent detector for biological chip |
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| CN100414288C true CN100414288C (en) | 2008-08-27 |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102353659B (en) * | 2011-06-09 | 2013-03-27 | 北京工业大学 | Detector for biochip fluorescent microspectrum and manufacture method thereof |
| CN105758834B (en) * | 2016-04-26 | 2018-11-27 | 福州大学 | A kind of biochip test method of induced with laser and CCD acquisition |
Citations (7)
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| US6197503B1 (en) * | 1997-11-26 | 2001-03-06 | Ut-Battelle, Llc | Integrated circuit biochip microsystem containing lens |
| CN1338830A (en) * | 2001-09-28 | 2002-03-06 | 清华大学 | Array-type integrated sensor based on weak-light electromechanical system for emitting or receiving laser |
| WO2002033385A2 (en) * | 2000-10-19 | 2002-04-25 | Motorola, Inc. | Biochip excitation and analysis structure |
| CN2501046Y (en) * | 2001-10-31 | 2002-07-17 | 东北大学 | Miniflow controlled chip analysis semiconductor laser induced fluorescence detector |
| JP2004257737A (en) * | 2003-02-24 | 2004-09-16 | Mitsui Eng & Shipbuild Co Ltd | Biochip reader |
| CN1563998A (en) * | 2004-04-16 | 2005-01-12 | 南京大学 | Microcurrent controlled chip detection combiner having general and integral and high efficiency radiation |
| CN2814401Y (en) * | 2005-07-27 | 2006-09-06 | 北京工业大学 | Millimeter micro laser induction fluorescent detector for biochip |
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2005
- 2005-07-27 CN CNB200510087115XA patent/CN100414288C/en not_active Expired - Fee Related
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6197503B1 (en) * | 1997-11-26 | 2001-03-06 | Ut-Battelle, Llc | Integrated circuit biochip microsystem containing lens |
| WO2002033385A2 (en) * | 2000-10-19 | 2002-04-25 | Motorola, Inc. | Biochip excitation and analysis structure |
| CN1338830A (en) * | 2001-09-28 | 2002-03-06 | 清华大学 | Array-type integrated sensor based on weak-light electromechanical system for emitting or receiving laser |
| CN2501046Y (en) * | 2001-10-31 | 2002-07-17 | 东北大学 | Miniflow controlled chip analysis semiconductor laser induced fluorescence detector |
| JP2004257737A (en) * | 2003-02-24 | 2004-09-16 | Mitsui Eng & Shipbuild Co Ltd | Biochip reader |
| CN1563998A (en) * | 2004-04-16 | 2005-01-12 | 南京大学 | Microcurrent controlled chip detection combiner having general and integral and high efficiency radiation |
| CN2814401Y (en) * | 2005-07-27 | 2006-09-06 | 北京工业大学 | Millimeter micro laser induction fluorescent detector for biochip |
Non-Patent Citations (2)
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| 半导体固体光源诱导荧光检测系统的研制. 杨丙成，谭峰，陈令新，关亚风.生命科学仪器，第1卷第2期. 2003 |
| 半导体固体光源诱导荧光检测系统的研制. 杨丙成，谭峰，陈令新，关亚风.生命科学仪器，第1卷第2期. 2003 * |
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