CN1616958A - Cell quantitative analysis method based on micro fluid control chip - Google Patents
Cell quantitative analysis method based on micro fluid control chip Download PDFInfo
- Publication number
- CN1616958A CN1616958A CN 200310105075 CN200310105075A CN1616958A CN 1616958 A CN1616958 A CN 1616958A CN 200310105075 CN200310105075 CN 200310105075 CN 200310105075 A CN200310105075 A CN 200310105075A CN 1616958 A CN1616958 A CN 1616958A
- Authority
- CN
- China
- Prior art keywords
- cell
- micro
- fluidic chip
- qualitative analysis
- pond
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The qualitative cell analysis method based on microfluid control chip features the single channel microfluid control chip as platform, the sample pond and the buffering pond on two ends of the channel and microscope coupled in any position to the channel. The analysis process includes: feeding cell liquid into the buffering pond and buffering liquid into the waste liquid pond and regulating the liquid surface height in the buffering pond to make cells inside the visual field still; applied voltage across the buffering pond and the waste liquid pond, recording with CCD on the microscope the directional shift speed of each cell and calculating the electrophoresis mobility of each cell; drawing the cell number- electrophoresis mobility curve of the tested cells; and comparing the curve with standard curves to obtain the qualitative cell analysis result. The method of the present invention has less interference and no need of separation or sorting of cell groups.
Description
Technical field:
The present invention relates to cell qualitative analysis technology, providing a kind of especially is the cell method for qualitative analysis of parameter with the electrophoretic mobility based on micro-fluidic chip.
Background technology:
Cytological classification is one of means of understanding the complex cell group.Is that it can realize pair cell group's Preliminary study without detachment process with cell sorting with separating different.Existing commercial instrument all needs pair cell to carry out sorting or separation and finishes cytological classification, does not still need the classifying instrument of cell separation to occur.
The cell electrophoresis mobility is that [Lin Kechun edits " biophysics " Beijing to one of parameter of characterize cells surface charge: People's Education Publishing House, 1981; 267-282], its measurement means commonly used comprises cell electrophoresis [Hashimoto, N., Fujita, s., Yokoyama, T., Ozawa, Y., Kingstsu, I., Kurosaka, D., Sabolovic, D., Schuett, W., Electrophoresis 1998,19,1227-1230.] [Liu Jian, Li Anli Chinese biological engineering in medicine journal 1990,9 (3), 161-191], Capillary Electrophoresis (CE) [Daniel W.Armstrong; MarcoGirod, Lingfeng He, Michael A.Rodriguez, Wei Wei, Jinjian Zheng and Edward S.Yeung.Anal Chem, 2002,74:5523], laser electrophoresis [Mao Huisheng, state's biomedical engineering newspaper 1994,13 (2) among strong, the Li An of blue indigo, horse recklessly, 171-174] etc.Comparatively speaking, cell electrophoresis instrument equipment is simple relatively, but owing to require vernier focusing in quiescent layer, is easy to generate big measuring error [Yan Zongyi, Shan Huixian.China's Hemorheology magazine 2001,11 (1), 1-4].CE measures the measurement that the cell electrophoresis mobility is difficult to accomplish individual cells.Sample introduction length, the effect of effect between the cell and cell and tube wall is bigger to the measuring accuracy influence.Though the laser electrophoresis apparatus is measured accurately, its apparatus expensive.The micro-fluidic chip technology that the nineties grows up has that analysis speed is fast, reagent consumption is little, easily be automated and advantage such as integrated, is used widely at the life science that comprises cell analysis.The microchannel has and is detection window everywhere, dark wide only be tens microns characteristics, can be coupled with microscope easily, realize the microelectrophoresis in the microchannel, and do not have quiescent layer, the reduction measuring error.But the report of research cell electrophoresis mobility seldom on the micro-fluidic chip platform, the earliest by Ichiki[Takanori Ichiki., TatekazuUjiie., Satomi Shinbashi., Tomoko Okuda., Electrophoresis 2002,23,2029-2034] etc. the people carry out, they investigated sheep red blood cell on quartz chip electrophoretic mobility and antibody to the influence of sheep red blood cell electrophoretic mobility.Comparing on the chip cell electrophoresis mobility measuring method with the laser electrophoresis with CE, to have equipment simple, the characteristics of flexible operation.No matter the method for above-mentioned any measurement electrophoretic mobility is not all mentioned its application in cytological classification.
Summary of the invention:
The object of the present invention is to provide a kind of is the cell qualitative classification analytical approach of parameter with the electrophoretic mobility based on micro-fluidic chip, and this method is disturbed little, need not the pair cell group separate or sorting, and instrument is simple, flexible operation.
The invention provides a kind of cell method for qualitative analysis, it is characterized in that based on micro-fluidic chip:
With single pass micro-fluidic chip is platform, and the passage two ends are respectively Buffer Pool and waste liquid pool, at arbitrary position coupling microscope of passage;
Process is:
Cell liquid is added in the Buffer Pool, and damping fluid joins in the waste liquid pool, regulates the Buffer Pool liquid level, and it is static that the field range inner cell is in;
On Buffer Pool and waste liquid pool, apply voltage, note the speed that each cell directional moves, and calculate the electrophoretic mobility (migration velocity under the mobility unit of the meaning field intensity) of each cell in view of the above by the electron-coupler (CCD) that is installed on the microscope;
With the electrophoretic mobility is horizontal ordinate, and the quantity of cell is the draw distribution spectral line of cell to be measured of ordinate;
The distribution spectral line of cell to be measured and the spectrum peak of standard spectral line are compared, obtain the classification qualitative results of cell.
In the cell method for qualitative analysis based on micro-fluidic chip provided by the invention, used cell concentration should be 10
3Individual/mL-10
8Between individual/mL.When cell concentration is lower than this scope, length consuming time.When being higher than this scope, because the chance that cell is in contact with one another and contacts with passage is big, statistical error increases.
In the cell method for qualitative analysis based on micro-fluidic chip provided by the invention, the electric field scope that applies depends on chip used volt-ampere characteristic, and the electric field intensity of electric field intensity when being no more than the volt-ampere curve off-straight is advisable.
In the cell method for qualitative analysis based on micro-fluidic chip provided by the invention, described chip material can be a polymkeric substance, glass, pottery or their complex.
In the cell method for qualitative analysis based on micro-fluidic chip provided by the invention, described channel width is between 20 microns to 2000 microns, between dark 30 microns to 200 microns.
In the cell method for qualitative analysis based on micro-fluidic chip provided by the invention, described cell can be the cell without any processing, also can be the cell that indicates various antibody or other molecules.
In addition, suitable equally for acellular charged particle (diameter is from several microns to tens microns) the present invention.
The microscopic cells electrophoresis apparatus that the inventive method is more traditional is subjected to littler method and disturbs, and more can reflect the character of cell or microparticle self really.The inventive method need not the pair cell group be separated or sorting.Used instrument is simple, flexible operation.
Description of drawings:
Fig. 1 is the migration photo of cell in passage;
Fig. 2 is the linear relation of cell migration speed and electric field intensity, and the slope of this straight line is the apparent electrophoretic mobility of cell;
Fig. 3 is the electrophoretic mobility three groups of HRD of RBC showed of classifying;
Fig. 4 be the morphological observation method normal-experimental group red blood cell photo;
Fig. 5 is morphological observation method cyhalothrin-experimental group red blood cell photo;
Fig. 6 is morphological observation method cyhalothrin-control group red blood cell photo.
Embodiment:
Get the 13uL sample and join in the buffer pool, because the difference in height of liquid level, liquid is full of each passage continuously; the damping fluid that in waste liquid pool, adds 13uL then; the careful liquid level of adjusting each liquid storage tank makes particle or cell keep static in passage, eliminates the influence of differential static pressure.In Buffer Pool, connect positive voltage subsequently, waste liquid pool ground connection, particle or cell are moved to negative pole by positive pole.Microscope imaging, the CCD record.CCD record result was 25 frame/seconds, and selected individual particle writes down its initial moment, preserves this moment picture and sees Fig. 1, writes down its reference position.Find same particle to stop constantly and final position with the frame progression, thereby calculate this particle speed.Each electric field writes down numbers of particles down greater than 30.
Fig. 3 is the unit field intensity velocity distribution and the normal distribution match thereof of all cells that records in the experiment.Three match peaks are corresponding with three groups of experimental cells.First group and the 3rd group of match peak realization " baseline separation ", second group " peak " is relatively poor with other two groups of " peak " " degree of separation ".The morphological observation of these three groups of cells the results are shown in Figure 4～6.Fig. 4,5 showed cell size is consistent, and haemoglobin is evenly distributed, and the two differentiating forms is very not remarkable.Fig. 6 can find out that then cell has dissolving trend.The classification of morphological observation and electrophoresis shows that all first group and second group of cytological classification conspicuousness are not strong, and both can replenish mutually, mutually checking.Explanation simultaneously, close at least with the result of electrophoretic mobility classification cell with morphologic observation cytological classification result, see Table 1.
Table 1 is three groups of carp erythrocyte electrophoresis mobilities.
| Group number | Group name | Electric osmose mobility cm 2/Vs | The apparent mobility cm of cell 2/Vs | The mobility cm of cell 2/Vs |
| ?NO.1 | Normally-experimental group | 1.8379E-04 | 7.00E-05 | ?1.1379E-04 |
| ?NO.2 | Cyhalothrin-experimental group | 1.71E-04 | ?1.279E-05 | |
| ?NO.3 | Cyhalothrin-control group | 2.69E-04 | ?-8.521E-05 |
Claims (6)
1, a kind of cell method for qualitative analysis based on micro-fluidic chip is characterized in that:
With single pass micro-fluidic chip is platform, and the passage two ends are respectively sample cell and Buffer Pool, at arbitrary position coupling microscope of passage;
Process is:
Cell liquid is added in the Buffer Pool, and damping fluid joins in the waste liquid pool, regulates the Buffer Pool liquid level, and it is static that the field range inner cell is in;
On Buffer Pool and waste liquid pool, apply voltage, note the speed that each cell directional moves by the electron-coupler (CCD) that is installed on the microscope, and calculate the electrophoretic mobility of each cell in view of the above;
With the electrophoretic mobility is horizontal ordinate, and the quantity of cell is the draw distribution spectral line of cell to be measured of ordinate;
The distribution spectral line of cell to be measured and the spectrum peak of standard spectral line are compared, obtain the classification qualitative results of cell.
2, according to the described cell method for qualitative analysis based on micro-fluidic chip of claim 1, it is characterized in that: used cell concentration should be 10
3Individual/mL-10
8Between individual/mL.
3, according to the described cell method for qualitative analysis of claim 1, it is characterized in that: the electric field intensity the when electric field intensity that applies is no more than chip used volt-ampere curve off-straight based on micro-fluidic chip.
4, according to the described cell method for qualitative analysis based on micro-fluidic chip of claim 1, it is characterized in that: described chip material is a polymkeric substance, glass, pottery or their complex.
5, according to the described cell method for qualitative analysis based on micro-fluidic chip of claim 1, it is characterized in that: described channel width is between 20 microns to 2000 microns, between dark 30 microns to 200 microns.
6, according to the described cell method for qualitative analysis based on micro-fluidic chip of claim 1, it is characterized in that: described cell is the cell without any processing, perhaps indicates the cell of various antibody or other molecules.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310105075 CN1616958A (en) | 2003-11-11 | 2003-11-11 | Cell quantitative analysis method based on micro fluid control chip |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310105075 CN1616958A (en) | 2003-11-11 | 2003-11-11 | Cell quantitative analysis method based on micro fluid control chip |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1616958A true CN1616958A (en) | 2005-05-18 |
Family
ID=34757176
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200310105075 Pending CN1616958A (en) | 2003-11-11 | 2003-11-11 | Cell quantitative analysis method based on micro fluid control chip |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1616958A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100489525C (en) * | 2006-06-27 | 2009-05-20 | 中国科学院力学研究所 | Cell organism microsystem for detecting cell surface marker and detection method thereof |
| CN101701895B (en) * | 2009-11-11 | 2011-04-13 | 华东理工大学 | Device and method capable of measuring liquid-liquid diffusion coefficient at different temperatures |
| CN101285083B (en) * | 2008-05-30 | 2011-06-29 | 华中科技大学 | Process for preparing patterned cellulosic by micro-fluidic chip |
| CN104122316A (en) * | 2014-06-30 | 2014-10-29 | 中国科学院化学研究所 | Method for simultaneously measuring mobility and dielectric mobility of particles by use of distributed direct-current electric field |
| WO2017028342A1 (en) * | 2015-08-14 | 2017-02-23 | 深圳大学 | Cell classification method based on light-induced dielectrophoresis technique |
| CN108603828A (en) * | 2015-11-30 | 2018-09-28 | 因塔生物公司 | Device and method for sample characterization |
-
2003
- 2003-11-11 CN CN 200310105075 patent/CN1616958A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100489525C (en) * | 2006-06-27 | 2009-05-20 | 中国科学院力学研究所 | Cell organism microsystem for detecting cell surface marker and detection method thereof |
| CN101285083B (en) * | 2008-05-30 | 2011-06-29 | 华中科技大学 | Process for preparing patterned cellulosic by micro-fluidic chip |
| CN101701895B (en) * | 2009-11-11 | 2011-04-13 | 华东理工大学 | Device and method capable of measuring liquid-liquid diffusion coefficient at different temperatures |
| CN104122316A (en) * | 2014-06-30 | 2014-10-29 | 中国科学院化学研究所 | Method for simultaneously measuring mobility and dielectric mobility of particles by use of distributed direct-current electric field |
| WO2017028342A1 (en) * | 2015-08-14 | 2017-02-23 | 深圳大学 | Cell classification method based on light-induced dielectrophoresis technique |
| CN108603828A (en) * | 2015-11-30 | 2018-09-28 | 因塔生物公司 | Device and method for sample characterization |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5578460A (en) | Electrophoretic method for the isolation and separation of microorganisms and cell populations | |
| Tibbe et al. | Optical tracking and detection of immunomagnetically selected and aligned cells | |
| US10578602B2 (en) | Disposable chamber for analyzing biologic fluids | |
| US10710074B2 (en) | Method for laser separation and characterization of particles and molecular species | |
| EP3045545B1 (en) | Method and apparatus for characterizing and counting particles, in particular biological particles | |
| Nakamura et al. | Separation of a breast cancer cell line from human blood using a quadrupole magnetic flow sorter | |
| US7250775B1 (en) | Microfluidic devices and methods based on measurements of electrical admittance | |
| US9110010B2 (en) | Electrical detection using confined fluids | |
| Vykoukal et al. | Dielectric characterization of complete mononuclear and polymorphonuclear blood cell subpopulations for label-free discrimination | |
| WO2005121767A1 (en) | Microfluidic device and analyzing/sorting device using the same | |
| Elitas et al. | Dielectrophoretic characterization and separation of monocytes and macrophages using 3D carbon‐electrodes | |
| CN106644900A (en) | Pulse impedance particle counting device based on non-uniform electric field and particle counting method | |
| EP3418721A1 (en) | A microfluidic chip | |
| Chen et al. | Direct characterization of motion-dependent parameters of sperm in a microfluidic device: proof of principle | |
| Zahedi Siani et al. | Dielectrophoretic separation of monocytes from cancer cells in a microfluidic chip using electrode pitch optimization | |
| CN1616958A (en) | Cell quantitative analysis method based on micro fluid control chip | |
| DE4124778A1 (en) | METHOD AND ARRANGEMENT FOR ANALYZING AGGLUTINATION REACTIONS | |
| CN206489050U (en) | A kind of impedance pulse grain count device based on inhomogeneous field | |
| CN108279202A (en) | A kind of adjustable grain count device and method of accuracy of detection | |
| CN202548092U (en) | Capillary electrophoresis detection system for relative content of glycosylated hemoglobin HbA1c | |
| CN111235021B (en) | Double-liquid-phase separation and detection device and method for circulating tumor cells in peripheral blood | |
| O’Connor | Flow cytometry versus fluorescence microscopy | |
| Tsukahara et al. | Dielectrophoresis of microbioparticles in water with planar and capillary quadrupole electrodes | |
| US11698357B2 (en) | Method for screening, isolating and purifying analytes | |
| Su | The development of an automated system for electrical characterization of cells using a novel force balance method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |