CN105719938A - Electrospray ionization device and application thereof - Google Patents
Electrospray ionization device and application thereof Download PDFInfo
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- CN105719938A CN105719938A CN201610080145.6A CN201610080145A CN105719938A CN 105719938 A CN105719938 A CN 105719938A CN 201610080145 A CN201610080145 A CN 201610080145A CN 105719938 A CN105719938 A CN 105719938A
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/02—Details
- H01J49/10—Ion sources; Ion guns
- H01J49/16—Ion sources; Ion guns using surface ionisation, e.g. field-, thermionic- or photo-emission
- H01J49/165—Electrospray ionisation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
Abstract
The invention relates to an electrospray ionization device. The electrospray ionization device comprises a glass substrate, a conductive clamp and a chamber, wherein the glass substrate has at least one pointed end, and is provided with a plurality of micro-channels; the conductive clamp is used for applying a high voltage and clamping the glass substrate; and the chamber is used for accommodating the glass substrate. The invention also relates to an application of the electrospray ionization device to cell-based drug analysis. Relatively high biocompatibility is provided through the glass substrate. The electrospray ionization device can be applied to cell-based qualitative and quantitative drug analysis and bioanalysis of cells after drug effects.
Description
Technical field
The present invention relates to the pharmaceutical analysis field based on cell, particularly relate to a kind of electro-spray ionization device and based on the application in the pharmaceutical analysis of cell.
Background technology
Pharmaceutical analysis based on cell plays an important role always in the development of biological medicine and pharmaceutical field.Traditional pharmaceutical analysis method based on cell needs to cultivate a large amount of cell, and could obtain sufficient data through complicated operation sequence.These traditional analysis methods often exist that reagent consumption is big, consuming time and the shortcoming of small throughput.
Mass spectrum is then that one comparatively quickly analyzes method, and the method can provide high sensitivity, high flux and molecular level other chemical information.As a kind of multi-functional analytical technology, mass spectrum can to the coupling such as electrophoresis, chromatograph to separate and to extract relevant ingredient, thus improving the sensitivity of pharmaceutical analysis, degree of accuracy and specific aim.Document 1, by Electrospray Mass Spectrometry and capillary electrophoresis coupling, can automatically analyze the haemachrome in single erythrocyte and α and the β substituent group in hemoglobin.But, sample for mass spectral analysis generally requires the off-line pretreatment first carrying out complexity, therefore, can only carry out departing from the off-line pharmaceutical analysis of cell micro-environment, and effectively pharmaceutical analysis requires that analysis condition should be as closely as possible to internal microenvironment, this causes that sample pre-treatments becomes a mass spectrum big obstacle for pharmaceutical analysis.
In order to overcome above-mentioned difficulties, people start to develop normality ionization massspectrum, to omit sample pre-treatments step, it is achieved analyze in its natural state and identify sample.Document 2 utilizes a capillary tube short, taper as receiving the sampling and spray tool upgraded, produces gas ion before mass detector, and the molecule that can be used for detecting in series of complex substrate is without sample pre-treatments.Document 3 then utilizes the sample spraying that puncture needle is made directly in tissue, can obtain accurate molecular information.Document 4 and document 5 then carry out electron spray with medical cotton stick and toothpick respectively, for pharmaceutical analysis.At this wherein, paper substrate electron spray is developing progressively the normality ioning method into a kind of maturation, for the analysis of complex sample.Electrolyte transports the tip of paper substrate by capillarity, forms ionspray after applying high electric field.Paper electrospray mass spectrometry, due to its shirtsleeve operation and the equipment that is easy to get, is usually used in the monitoring of the biomolecule detection in cell culture medium, biological fluid analysis and food safety.Cellulose chromatography paper and nitrocellulose filter are conventional paper substrates.But, these paper substrates make cell be difficult to adhere to thereon because of its poor biocompatibility, cannot realize again because of its opacity the optical instrument of cell is monitored in real time, thus limiting it based on the application in the research of cell.Accordingly, it is capable to the apparatus and method being enough in the normality ionizing of the pharmaceutical analysis based on cell urgently develop.
Correlation technique document
Document 1:IntegratedMicrofluidicDeviceforAutomatedSingleCellAnal ysisUsingElectrophoreticSeparationandElectrosprayIonizat ionMassSpectrometry (MellorsJ.S.；JorabchiK.；SmithL.M.；RamseyJ.M.Anal.Chem., 2010,82,967-973).
Document 2:CapillaryAction-SupportedContactlessAtmosphericPressur eIonizationfortheCombinedSamplingandMassSpectrometricAna lysisofBiomolecules (HsiehC.；ChangC.；UrbanP.L.；ChenY.Anal.Chem., 2011,83,2866-2869).
Document 3:Biologicaltissuediagnosticsusingneedlebiopsyandsprayio nizationmassspectrometry (LiuJ.；CooksR.G.；OuyangZ.Anal.Chem., 2011,83,9221-9225).
Document 4:Directdruganalysisfromoralfluidusingmedicalswabtouchsp raymassspectrometry (PirroV.；JarmuschA.K.；VincentiM.；CooksR.G.Anal.Chim.Acta, 2015,861,47-54).
Document 5:Internalstandardmassspectrumfingerprint:Anovelstrategy forrapidassessingthequalityofShuang-Huang-Lianoralliquid usingwooden-tipelectrosprayionizationmassspectrometry (YangY.；DengJ.Anal.Chim.Acta, 2014,837,83-92).
Summary of the invention
In view of above-mentioned technical problem, it is an object of the invention to provide the electro-spray ionization device that a kind of normality ionization massspectrum for the pharmaceutical analysis based on cell detects, higher biocompatibility is provided by substrate of glass, can be used for the qualitative and quantitative analysis of cell Chinese medicine, it may also be used for the bioanalysis of the cell after medicine effect.
An embodiment of the invention is in that to provide a kind of electro-spray ionization device, including:
There is the substrate of glass at least one tip, be provided with multiple microchannel；
For applying high voltage and clamping the conductive clamp of described substrate of glass；And
For placing the chamber of described substrate of glass.
The present inventor it have been investigated that, by higher biocompatibility can be provided based on the electro-spray ionization device of substrate of glass, thus being easily achieved cell adherent and growth thereon, simultaneously because the transparency of substrate of glass, it is also easy to realize the real-time monitoring to cell.On the other hand, by arranging multiple microchannel on the glass substrate, due to the surface tension effects of microchannel, will not be mutually mixed between cell, co-culturing of various kinds of cell can be realized, thus being conducive to cell medicine effect under co-culturing condition is carried out qualitative or detection by quantitative.
One of the present invention preferred embodiment in, the plurality of microchannel is arranged on the glass substrate by photo etched mask method.
According to the present invention, after the negative optical cement of spin coating on the glass substrate, baking, by the mask that is decorated with microchannel pattern, (channel portion is light-transmissive film, remainder is light tight) cover in the substrate of glass scribbling negative optical cement, litho machine carries out uv-exposure 3 times, each 30s, so that the negative light adhesive curing covered by photo mask；Then take mask off and develop with developer solution, to remove uncured negative optical cement；Drying up with nitrogen, in substrate of glass, because of solidification, substrate of glass is separated into multiple microchannel by not removed negative optical cement again, namely obtains being provided with the substrate of glass of microchannel.
The present invention another preferred embodiment in, the width of the plurality of microchannel is 1-3mm, and the degree of depth is 10-50 μm, and number is 2-5.
The present invention another preferred embodiment in, described substrate of glass is for being cut into acute triangle, it is preferred to the coverslip of isosceles acute triangle, and the length of side is respectively less than 1cm, and thickness is 5-20 μm.
The present invention another preferred embodiment in, described electro-spray ionization device includes sheet polymer material, and described chamber is formed on described sheet polymer material, and the shape of the shape of described chamber and described substrate of glass adapts.
Shape according to the present invention, the shape of described chamber and described substrate of glass adapts, and represents that described chamber is same or similar with the shape of described substrate of glass, and the area of described chamber is slightly larger than described substrate of glass.
In a specific embodiment of the present invention, described chamber is formed as follows: prepare the onesize sheet polymer material with shape of two panels, a cavity adapted with the shape of described substrate of glass is dug out on a piece of wherein, it is layered on another sheet polymer material, thus constituting chamber.
The present invention another preferred embodiment in, described sheet polymer material is organosilicon polymer, it is preferred to polydimethylsiloxane (PDMS).
Another embodiment of the invention is in that a kind of method providing Mass Spectrometer Method utilizing above-mentioned electro-spray ionization device to carry out Intracellular drug absorption, including:
It is injected separately into cell suspension overnight incubation in multiple microchannels of substrate of glass；
Medicine is adopted to act on the cell to be measured of overnight incubation；
For being detected by mass spectrograph through the medicine of pharmaceutically-active Cell uptake to be measured.
One of the present invention preferred embodiment in, the time adopting the cell to be measured that medicine acts on overnight incubation is 24h.
The present invention another preferred embodiment in, carry out detection by mass spectrograph to include: clamp before substrate of glass is positioned over mass spectrograph with conductive clamp, by substrate of glass tip just to mass spectrometric injection port, high-tension electricity is applied by conductive clamp, to dripping organic solvent in the microchannel at cell place to be measured, detect after producing electron spray.
Yet further embodiment of the invention is in that to provide a kind of method utilizing above-mentioned electro-spray ionization device to carry out apoptosis rate detection, including:
It is injected separately into cell suspension overnight incubation in multiple microchannels of substrate of glass；
Medicine is adopted to act on the cell to be measured of overnight incubation；
Dye to through pharmaceutically-active cell to be measured with fluorescent probe, and analyze apoptosis situation.
The further embodiment of the present invention is in that to provide above-mentioned electro-spray ionization device based on the application in the pharmaceutical analysis of cell.
According to electro-spray ionization device provided by the present invention, higher biocompatibility is provided by substrate of glass, co-culturing of various kinds of cell is realized by microchannel, can be widely applied for the pharmaceutical analysis in the cell after medicine effect and bioanalysis, particularly in in cell pharmaceutical analysis under co-culturing state and bioanalysis, there is obvious advantage.Method according to Mass Spectrometer Method provided by the present invention, it is possible to realize the normality ionizing of the pharmaceutical analysis based on cell, is conducive to improving degree of accuracy and the repeatability of pharmaceutical analysis.
Accompanying drawing explanation
Fig. 1 is shown that the structural representation of the glass base electro-spray ionization device according to the present invention and mass spectrometry, in FIG, and 1-conductive clamp；2-substrate of glass；3-sheet polymer material；4-chamber；5-organic solvent；6-electron spray；7-mass spectrograph injection port.
The schematic diagram carrying out three kinds of co-culture of cells of HepG2, Caco-2 and MCF-7 according to embodiments of the invention 2 that Fig. 2 shows.
Fig. 3 is shown that carrying out the laser co-focusing microphotograph of three kinds of co-culture of cells of HepG2, Caco-2 and MCF-7 according to embodiments of the invention 2.
Fig. 4 is shown that carrying out three kinds of cell survival rate analysis charts of co-culture of cells 1-3 days of HepG2, Caco-2 and MCF-7 according to embodiments of the invention 2, is shown that the result of three groups of parallel laboratory tests in figure.
Fig. 5 is shown that the qualitative Mass Spectrometer Method figure according to the embodiments of the invention 3 systemic CPA of MCF-7 cell to co-culturing, and the illustration in the upper right corner is molecular formula and the molecular weight of CPA.
Fig. 6 is shown that according to the embodiments of the invention 3 qualitative Mass Spectrometer Method figure to single systemic CPA of MCF-7 cell cultivated.
Fig. 7 is shown that according to embodiments of the invention 4 with isotope CPA-d4For the mass spectrum quantitative measurement standard curve that interior mark does.
Fig. 8 is shown that, according to the result of CPA absorbtivity in embodiments of the invention 4 detection by quantitative MCF-7 cell, being shown that the result of three groups of parallel laboratory tests in figure, and in figure, * indicates marked difference.
Fig. 9 is shown that being shown that in Apoptosis assay figure, the figure after the MCF-7 cell co-cultured and list is cultivated being carried out CPA medicine effect according to embodiments of the invention 5 result of three groups of parallel laboratory tests, and in figure, * indicates marked difference.
Figure 10 is shown that being shown that in Apoptosis assay figure, the figure after the MCF-7 cell co-cultured and list is cultivated being carried out Dox medicine effect according to embodiments of the invention 5 result of three groups of parallel laboratory tests, and in figure, * indicates marked difference.
Detailed description of the invention
The present invention is described in detail by the following examples, but protection scope of the present invention is not limited to the description below.If without specified otherwise, it it is experimentally normal experiment method；The reagent used and material etc. all can pass through to be either commercially available.
The manufacture method of embodiment 1 electro-spray ionization device and structure thereof
(making of substrate of glass)
The coverslip diamant that thickness is 20 μm is cut into waist length is 0.8cm, drift angle is the isosceles triangle of 45 °, clean, dry after, the negative optical cement of SU-82015 being 30 μm with the rotating speed of 1000rpm in surface spin coating a layer thickness with sol evenning machine, 20min is toasted at 65 DEG C, by being decorated with microchannel pattern, (width of described microchannel is 2mm, number is 3) mask cover on slide, after uv-exposure, development, nitrogen dry up, namely obtain being provided with 3 width to be 2mm, the degree of depth be the substrate of glass of 30 μm of microchannels.
(making of PDMS chamber)
It is poured on silicon chip after PDMS prepolymer is mixed with mass ratio 10:1 with initiator, seals with wide adhesive tape in advance around silicon chip, it is prevented that PDMS prepolymer spills.By placing 2h after PDMS prepolymer bubble removing at 75 DEG C, carry out polyreaction.PDMS sheet after polymerization is carefully taken off, cuts into the PDMS sheet of 2cm × 2cm × 5mm.Make one piece of an equal amount of PDMS sheet by same method again, and dig out an isosceles triangle cavity (the waist length in this isosceles triangle cavity is 1.2cm, and drift angle is 45 °) thereon.By digging the PDMS sheet in isosceles triangle cavity and not having empty PDMS sheet stacked together, define the PDMS chamber that can hold above-mentioned substrate of glass, substrate of glass is put into wherein.
Conductive clamp: prepare conductive metal clip, standby.
Embodiment 2 cell co-culturing in microchannel
1) by the human liver tumor cell HepG2 covered with in 60mm culture dish, people clones colon adenocarcinoma cell Caco-2 and human breast cancer cell line Bcap-37 cell digests respectively, and respectively with culture medium dilution be 106mL-1Cell suspension.The cell suspension of three kinds of cells is respectively taken 10 μ L and is separately added in three microchannels of the substrate of glass being placed in PDMS chamber (as shown in Figure 2), after cell attachment, adding some new culture medium, the substrate of glass being placed in PDMS chamber puts into cell culture incubator, overnight incubation.
2) from incubator, substrate of glass is taken out, with the fluorescent dye that CellTrackerTMGreenCMFDA, CellTrackerTMDeepRed and CellTrackerTMViolet are three kinds different, HepG2, Caco-2 and MCF-7 cell is dyeed respectively, being placed under laser confocal microscope to observe, result is as shown in Figure 3.It can be seen that due to the surface tension effects of microchannel, three kinds of cells grow in respective microchannel, will not be mutually mixed between cell.
3) according to step 1) identical mode by tri-kinds of cell overnight incubation of HepG2, Caco-2 and MCF-7, change culture medium every day, standby.Cell survival rate is detected every day with life or death agent box (Calcein-AM/EthD-1).As shown in Figure 4, cell all keeps normal in first three day activity cultivated to result, still reaches more than 85% at the 3rd day cell survival rate.
The qualitative Mass Spectrometer Method that embodiment 3 Intracellular drug absorbs
Step 1 according to embodiment 2) identical mode co-cultures HepG2, Caco-2 and MCF-7 cell, after overnight incubation, it is placed on the substrate of glass in PDMS chamber to take out from incubator, dropping cyclophosphamide solution is (hereinafter referred to as CPA, concentration is 10mmol/L, i.e. 10mM, and consumption is 10 μ L), then put back in incubator, cultivate 24h.
Substrate of glass all to cultivate MCF-7 cell (i.e. single MCF-7 cell cultivated) in three microchannels is identical with the MCF-7 cell co-cultured as control sample, its training method and medicine model of action.
From cell culture incubator, take out the substrate of glass being placed in PDMS chamber, substrate of glass is picked up, remain in the complex matrices of surface of glass slide with distilled water flushing for three times with removing.After substrate of glass is dried naturally, clip before being positioned over Shimadzu LCMS-2010A mass spectrograph with conductive clamp, make the tip (45° angle place) of substrate of glass just to mass spectrograph injection port, by conductive clamp, substrate of glass applied high-tension electricity (～4.5kV), in the microchannel at MCF-7 cell place, drip the isopropanol as organic solvent simultaneously, produce electron spray immediately, collect electrospray ionization with mass spectrograph and carry out scanning analysis.The mass-to-charge ratio (m/z) of scanning ranges for 100 to 500, detects with positive ion mode.Control sample is also scanned analyzing in the manner described above.
As it can be seen in figures 5 and 6, have [CPA+H] in all mass spectruies+Peak (m/z is 262), [CPA+Na]+Peak (m/z is 284) and [CPA+K]+Peak (m/z is 300).These typical mass spectra peaks show that CPA has absorption in the MCF-7 cell singly cultivated and co-culture.
The quantitative mass spectral detection that embodiment 4 Intracellular drug absorbs
Stable isotope CPA-d with CPA4For interior mark, employing Internal standard carries out mass spectrum quantitative analysis.Specifically, make 5 sheet glass substrates according to the method identical with embodiment 1, by the CPA-d of 5 μ L4Culture medium solution (10mmol/L) be separately added in the culture medium solution (5,10,20,40 and 60mmol/L) of CPA of 10 μ L series concentration, be delivered to respectively in the center-aisle of 5 sheet glass substrates.After its volatilization is dry, it is sequentially carried out scanning of the mass spectrum according to the mode identical with embodiment 3.With [CPA+Na]+Peak intensity and [CPA-d4+Na]+The ratio of peak intensity is relative peak intensities, with relative peak intensities for vertical coordinate, makes standard curve with CPA concentration for abscissa.As it is shown in fig. 7, standard curve shows well linear (R2=0.9981), linear equation is y=0.0652x+0.3256.Similar structure between CPA and isotope thereof effectively attenuates the impact of complex matrices.
Step 1 according to embodiment 2) identical mode co-cultures HepG2, Caco-2 and MCF-7 cell, after overnight incubation, it is placed on the substrate of glass in PDMS chamber to take out from incubator, the CPA solution (consumption 10 μ L) of dropping 10mmol/L, then put back in incubator, cultivate 24h.Identical with the MCF-7 cell co-cultured as control sample, its training method and medicine model of action using single MCF-7 cell cultivated.
From cell culture incubator, take out the substrate of glass being placed in PDMS chamber, substrate of glass is picked up, remain in the complex matrices of surface of glass slide with distilled water flushing for three times with removing.After substrate of glass is dried naturally, carry out scanning of the mass spectrum according to the mode identical with embodiment 3.Shown in the data obtained such as table 1 and Fig. 8.
As shown in table 1 and Fig. 8, different in the amount of the systemic CPA of MCF-7 cell singly cultivating and co-culturing, the MCF-7 cell co-cultured is bigger than normal to the absorbtivity of CPA.Average absorption amount respectively 2.33mmol/L and the 0.07mmol/L of CPA in the MCF-7 cell co-cultured and singly cultivate.Very well, RSDs value is respectively less than 10% to the repeatability of experiment.
The relative standard deviation (RSDs) of CPA absorbtivity in table 1 cell
The drug-induced Apoptosis assay of embodiment 5
CPA and amycin (Dox) act on as two kinds of model drugs and co-culture with on single MCF-7 cell cultivated, and both anticarcinogens can cause apoptosis.By CPA storing solution (100mmol/L) and Dox storing solution (10mg/mL) respectively with cell culture medium stepwise dilution to 10mmol/L and 2 μ g/mL.Step 1 according to embodiment 2) identical mode co-cultures HepG2, Caco-2 and MCF-7 cell, after overnight incubation, act on cell 24h according to the mode identical with embodiment 3 or 4 10 μ LCPA solution (10mmol/L) or 10 μ LDox solution (μ g/mL) respectively.Identical with the MCF-7 cell co-cultured as control sample, its training method and medicine model of action using single cultivation MCF-7 cell.
After 24 hours, from incubator, take out substrate of glass, with fluorescent probe Hoechst33342 to cell dyeing, take pictures with fluorescence microscope (LeicaDMI4000B), and analyze apoptosis situation with software I mage-ProPlus6.0, shown in result such as table 2 and Fig. 9, Figure 10.
Under same drug level, the MCF-7 cells show singly cultivated and co-culture goes out different apoptosis rates.Under CPA effect, the MCF-7 cell co-cultured goes out higher apoptosis rate than single MCF-7 cells show cultivated, and under Dox effect, single MCF-7 cell cultivated goes out higher apoptosis rate than the MCF-7 cells show co-cultured.Inventor speculates, this is owing to CPA is a kind of prodrug, only just can cause the apoptosis of breast cancer cell after liver metabolism, so going out higher apoptosis rate with the MCF-7 cells show of HepG2 co-culture of cells.Dox then can not pass through Caco-2 cell, and therefore, the MCF-7 apoptosis rate co-cultured declines on the contrary.Experiment shows good repeatability, and relative standard deviation value (RSDs) is respectively less than 10%.The apoptosis rate more caused greatly due to the absorbtivity of CPA can be more high.Therefore, the average absorption amount of CPA in the cell of Mass Spectrometer Method the difference of the MCF-7 iuntercellular apoptosis rate singly cultivated and co-culture is explained well.
The relative standard deviation (RSDs) of the drug-induced apoptosis rate of table 2
It should be noted that, embodiment described above is only used for explaining the present invention, it is not intended that any limitation of the invention.By referring to exemplary embodiments, invention has been described, it should be appreciated that word wherein used is descriptive and explanatory vocabulary, rather than limited vocabulary.Within the scope of the claims the present invention can be modified by regulation, and in without departing substantially from scope and spirit of the present invention, the present invention be revised.Although the present invention described in it relates to specific method, material and embodiment, it is not intended that the present invention is limited to wherein disclosed particular case, on the contrary, the present invention can be extended to other all methods and applications with identical function.
Claims (10)
1. an electro-spray ionization device, including:
There is the substrate of glass at least one tip, be provided with multiple microchannel；
For applying high voltage and clamping the conductive clamp of described substrate of glass；And
For placing the chamber of described substrate of glass.
2. electro-spray ionization device according to claim 1, it is characterised in that the plurality of microchannel is arranged on the glass substrate by photo etched mask method.
3. electro-spray ionization device according to claim 1 and 2, it is characterised in that the width of the plurality of microchannel is 1-3mm, and the degree of depth is 10-50 μm, number is 2-5.
4. the electro-spray ionization device according to any one of claim 1-3, it is characterised in that described substrate of glass is for being cut into acute triangle, it is preferred to the coverslip of isosceles acute triangle, and the length of side is respectively less than 1cm, and thickness is 5-20 μm.
5. the electro-spray ionization device according to any one of claim 1-4, it is characterized in that, described electro-spray ionization device also includes sheet polymer material, and described chamber is formed on described sheet polymer material, and the shape of the shape of described chamber and described substrate of glass adapts.
6. electro-spray ionization device according to claim 5, it is characterised in that described sheet polymer material is organosilicon polymer, it is preferred to polydimethylsiloxane.
7. utilize the electro-spray ionization device according to any one of claim 1-6 to carry out the method for Mass Spectrometer Method for drug absorption, including:
It is injected separately into cell suspension overnight incubation in multiple microchannels of substrate of glass；
Medicine is adopted to act on the cell to be measured of overnight incubation；
For being detected by mass spectrograph through the medicine of pharmaceutically-active Cell uptake to be measured.
8. the method for Mass Spectrometer Method according to claim 7, it is characterized in that, carry out detection by mass spectrograph to include: clamp before substrate of glass is positioned over mass spectrograph with conductive clamp, by substrate of glass tip just to mass spectrograph injection port, high-tension electricity is applied by conductive clamp, to dripping organic solvent in the microchannel at cell place to be measured, detect after producing electron spray.
9. utilize the method that the electro-spray ionization device according to any one of claim 1-6 carries out apoptosis rate detection, including:
It is injected separately into cell suspension overnight incubation in multiple microchannels of substrate of glass；
Medicine is adopted to act on the cell to be measured of overnight incubation；
Dye to through pharmaceutically-active cell to be measured with fluorescent probe, and analyze apoptosis situation.
10. the electro-spray ionization device described in any one of claim 1-6 is based on the application in the pharmaceutical analysis of cell.
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| US20050048581A1 (en) * | 2003-08-25 | 2005-03-03 | Chiu Daniel T. | Method and device for biochemical detection and analysis of subcellular compartments from a single cell |
| CN101643701A (en) * | 2009-07-23 | 2010-02-10 | 清华大学 | Cell sorter micro-fluidic chip based on immunomagnetic separation technology and application thereof in aspect of enrichment of rare cells |
| CN102590322A (en) * | 2012-01-31 | 2012-07-18 | 复旦大学 | Liquid drop microsystem applied to proteomic research |
| CN103226127A (en) * | 2013-03-27 | 2013-07-31 | 清华大学 | Multi-channel micro-fluidic chip and mass spectrum combined device |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050048581A1 (en) * | 2003-08-25 | 2005-03-03 | Chiu Daniel T. | Method and device for biochemical detection and analysis of subcellular compartments from a single cell |
| CN101643701A (en) * | 2009-07-23 | 2010-02-10 | 清华大学 | Cell sorter micro-fluidic chip based on immunomagnetic separation technology and application thereof in aspect of enrichment of rare cells |
| CN102590322A (en) * | 2012-01-31 | 2012-07-18 | 复旦大学 | Liquid drop microsystem applied to proteomic research |
| CN103226127A (en) * | 2013-03-27 | 2013-07-31 | 清华大学 | Multi-channel micro-fluidic chip and mass spectrum combined device |
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