CN104280329A - Micro-fluidic multicolor fluorescence cell counter - Google Patents
Micro-fluidic multicolor fluorescence cell counter Download PDFInfo
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Abstract
The invention relates to a micro-fluidic multicolor fluorescence cell counter. The micro-fluidic multicolor fluorescence cell counter comprises a micro-fluidic chip system, a fluorescent light source system and an analyzing and counting system, wherein the micro-fluidic chip system comprises a micro-fluidic chip (3), a computer-controlled controller (18) and a pipeline (20); a fluorescence imaging system comprises a laser A (9), a laser B (12), a fluorescence detection objective lens (8) and a signal processing vidicon (17); the analyzing and counting system comprises a computer (19). The micro-fluidic multicolor fluorescence cell counter is combined with a magnetic cell sorting technology and an immunocytochemical staining technology and is applied to detection of peripheral blood circulation cancer cells of colon cancer; the micro-fluidic analyzing and counting system is used for analyzing and counting tumor cells; the cells are counted; the cellular morphology analysis is achieved.
Description
Technical field
The invention belongs to FCM analysis field, particularly a kind of multicolor fluorescence cell counter based on micro-fluidic chip.
Background technology
Circulating tumor cell (CTCs) refers to and to spin off from primary tumor and to enter sanguimotor cell, and these cells have other position in body and adhere to and form the potential of new cancer metastasis.CTC is found in kinds cancer patient body, it has the biological property of primary tumor cell, can detect in the peripheral blood of the shallow table of body, be considered to have important using value in reflection cancer grade malignancy and prognosis, its number quantitative statistics likely becomes the important indicator judging cancer metastasis, recurrence, prognosis.Therefore, how efficiently, carry out that analysis of accounts become based on circulating tumor cell to the CTCs obtained fast and accurately is one of the key issue of main cancer diagnosis system research and development and application.
The method of general acquisition CTC uses biomolecular to catch the CTC in blood, after Magneto separate, carries out labeled analysis to the circulating tumor cell of catching.Its ultimate principle uses specific fluorescent dye to mark target cell, utilizes the fluorescence excitation light source in fluorescent microscope to excite, the fluorescence signal of static state or dynamic cellular after detection fluorescence labeling, and carry out statistical study.General fluorecyte calculating instrument can detect and count cycle tumour cell automatically, although these instruments have power, it also exists some areas for improvement.First, the relevant device of the photofluorometer counting apparatus in instrument need with CTCs Magneto separate acquisition equipment with the use of, this adds the cost of instrument itself and corresponding assembly undoubtedly, makes its more difficult popularization and application; The second, the biomolecular of catching CTC only adopts single anti-EpCAM antibody modification, and the unicity of biomolecular has had a strong impact on its capture rate to the tumour cell of complexity.3rd, the photofluorometer counting apparatus in instrument needs the number of cells by artificial counting fluorescent dye, and artificial counting not only adds the time of sample analysis, and reduces stability and the consistance of counting to a certain extent.Therefore, market and clinical cheap in the urgent need to one, has independent research advantage, and can well flexible Application in ordinary consumption colony CTCs fluorescence analysis and pass judgment on device.
Microfluidic chip technology is the subject grown up the nineties in 20th century, refers to the science and technology of fluid in the small network channel of operation (5-500 micron).Micro-fluidic chip is the micro device based on extensive parallel processing biological information molecule principle, microlitre (μ L) can be transmitted to receiving liter (nL) the even fluid of skin liter (pL) magnitude, and some steps of biochemical reaction comprised analysis, washing, detection etc. be integrated on one or a few micro-fluid chip, have that information flux is large, robotization, systematized feature.Micro-fluidic chip has integrability, the feature of easily automation mechanized operation makes it have much potential advantage compared with routine bioanalytical, has increasing application in bio-medical analysis field.Instrument such as PCR at present based on micro-fluidic chip, protein crystal instrument, DNA tests instrument is all come out one after another.
Summary of the invention
Invent technical matters to be solved
Technical matters to be solved by this invention provides a kind of instrument quick and precisely counted of fluorescence labeling Posterior circle tumour cell, fill up in conjunction with microfluidic chip technology and immunocytochemical technique, carry out the technological gap of circulating tumor cell determination and analysis accurately.
Technical scheme
One of technical scheme provided by the invention is a kind of micro-fluidic multicolor fluorescence cell counter, comprises micro-fluidic chip system, fluorescence light source system and analysis meter number system; Described micro-fluidic chip system comprises micro-fluidic chip, computer-controlled controller and pipeline; Described fluorescence light source system comprises two kinds or more laser instruments, as laser instrument A and laser instrument B, the object lens of fluorescence detection and the video camera of signal transacting; Described analysis meter number system comprises computing machine.
One of optimal way of above-mentioned technical scheme is, described micro-fluidic chip system also alternatively containing, objective table, motor, one of guide rail and screw mandrel or more.
Two of the optimal way of above-mentioned technical scheme is, described fluoroscopic imaging systems is also alternatively containing one of lighting source, condenser, object lens, beam expanding lens A, dichroic mirror 1, beam expanding lens B, dichroic mirror 2, optical filter runner or more.
Three of the optimal way of above-mentioned technical scheme is that described object lens are 40 times of Liars preferably, the described preferred 405nm of laser instrument A, the described preferred 488nm of laser instrument B.
Four of the optimal way of above-mentioned technical scheme is that the material of described micro-fluidic chip is silicone; There is three-dimensional structure; Chip thickness is less than 50 microns; In chip, sample capacity is less than 10 microlitres; Chip surface adopts EpCAM antibody modification.
Five of the optimal way of above-mentioned technical scheme is that described fluorescence light source system establishes two kinds of fluorescence excitation light sources; The cell that described laser instrument (A) fluorescent exciting source excitation is dyeed by 4', 6-diamidino-2-phenylindone (DAPI), produces blue-fluorescence; The cell that described laser instrument (B) fluorescent exciting source excitation is dyeed by fluorescein isothiocynate (FITC) and phycoerythrin (PE), produces green fluorescence and ruddiness fluorescence respectively.
Two of technical scheme provided by the invention is a kind of preparation method of micro-fluidic multicolor fluorescence cell counter, comprises and prepares micro-fluidic chip with following steps,
1) spin coating photoresist is in silicon chip surface, and solidification, develops after uv-exposure, makes micro-fluidic chip template;
2) silicone (PDMS) is 5:1 mixing by A:B, degassed in a vacuum, pours into and is equipped with in the double dish of mould, the thickness of falling glue 5 millimeters, be separated by PDMS after 80 DEG C of 60min that are heating and curing with mould;
3) punch, upper and lower two-layer PDMS is aimed at, bonding; 80 DEG C are toasted 1 hour.
Three of technical scheme provided by the invention is counting and the graphical analysis that above-mentioned cell counter is used for circulating tumor cell in blood.
Four of technical scheme provided by the invention is that above-mentioned cell counter is used for colorectal cancer circulating tumor cell counting.
Five of technical scheme provided by the invention is the application of above-mentioned cell counter, comprises and carries out circulating tumor cell Magneto separate and qualification in advance; Or, be included in micro-fluidic chip and carry out cellular morphology and fluorescence labeling state analysis.
Described micro-fluidic multicolor fluorescence cell counter is used for counting the circulating tumor cell in sample and analyzing;
Described fluorescence light source system is for carrying out fluorescence excitation to the tumour cell on micro-fluidic chip, taking pictures and cell count; Comprise a light supply apparatus near micro-fluidic chip, if three kinds of fluorescence excitation light sources, DAPI blue light, FITC green glow and PE ruddiness; The fluorescence detection device of a close described micro-fluidic chip and described light supply apparatus; A signal processor be connected with fluorescence detection device.
Described analysis meter number system comprises software and computer control module, produces data for the collection of fluoroscopic image and the Auto-counting of tumour cell and automatic analysis.
Multicolor fluorescence cell counter of the present invention is provided with computer interface, is controlled analysis meter number system by computing machine.
Tumour cell fluorescence detector critical optical part of the present invention can select omnipotent infinity correcting optical system.Light is by becoming parallel beam after the chromatic aberration correction object lens of infinity, and this parallel beam is imaged onto CCD sensitive chip through Guan Jing.Can optical accessory be added and not affect total magnification between object lens and Guan Jing.
The application of micro-fluidic multicolor fluorescence cell counter of the present invention comprises counting and the graphical analysis of circulating tumor cell in blood, need coordinate corresponding circulating tumor cell Magneto separate and identification systems during use; Also comprise the cellular morphology in micro-fluidic chip and fluorescence labeling state analysis in addition.
Beneficial effect
1) the present invention carries out programming count in conjunction with microfluidic chip technology, immunocytochemical technique and the multicolor fluorescence analysis technology number to cell particularly circulating tumor cell, the microscopic pattern of cell is carried out to collection and the analysis of image simultaneously, and can realize further the analysis of circulating tumor cell in molecule and gene, result reliability is high;
2) on the other hand, utilize analysis meter counting apparatus to carry out circulating tumor cell and detect and counting, reduce handling time and handling procedure.
3) the immune magnetic microsphere technology comprised with the matching used cell capturing system of micro-fluidic multicolor fluorescence cell counter of the present invention is based on immunology, utilizes the various microballoons being coated with immunologic active material to carry out the new technology of immunology or other Biological Detection; Technique has been widely used every field owing to having quick, easy, separation purity advantages of higher and has obtained huge progress.
Micro-fluidic chip involved in the present invention has following characteristics:
A. utilize micro-fluidic chip to the accurate manipulation ability of cell in micro-meter scale fluid, precise manipulation can be carried out to the CTC cell adding chip, by the Exact Design of micro-fluidic chip, realize CTC monolayer in chip;
B. the three-dimensional structure modeling of micro-fluidic chip, not only realize monolayer in chip, and maintain the structural strength (in chip bar shaped skeleton structure) of chip, design special marking simultaneously, facilitate the accurate positioning analysis CTC cell of multicolor fluorescence analysis instrument.
C. in conjunction with multicolor fluorescence analysis instrument, fluorescence analysis and the quick counter of individual cells is realized.
Accompanying drawing explanation
Figure l is the workflow schematic diagram of patent of the present invention.Legend: lighting source (1), condenser (2), micro-fluidic chip (3), objective table (4), motor (5), guide rail (6), screw mandrel (7), object lens (8) (preferably 40 times), laser instrument A (9), (preferred 405nm) beam expanding lens A (10), dichroic mirror 1 (11), laser instrument B (12) (preferred 488nm), beam expanding lens B (13), dichroic mirror 2 (14), optical filter runner (15), Guan Jing (16), video camera (17), controller (18), computing machine (19), pipeline (20),
Fig. 2 is the structural representation of micro-fluidic chip (3);
Fig. 3 be micro-fluidic chip (3) upper read carry out immunostaining after cell fluorescence MIcrosope image.
Embodiment
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.In addition should be understood that those skilled in the art can make various changes or modifications the present invention, and these equivalent form of values fall within the application's appended claims limited range equally after the content of having read the present invention's instruction.
Embodiment 1
(1) the catching and fluorescence labeling of colorectal cancer CTC
Colorectal cancer cell (Tumor Hispital Attached to Fudan Univ, 10 parts, every part of 5.0mL) in peripheral blood CTC high-purity separation kit (Bai Huikang bio tech ltd, Shanghai) human peripheral blood is adopted to carry out catching and fluorescence labeling.Two processes are divided into the analog capture experiment of colorectal cancer cell, first by colorectal cancer HT29 cell and other normal cell (293T cell, lymphocyte and macrophage (American Type Culture collection warehousing (ATCC))) blended checking peripheral blood CTC high-purity separation kit is to the separation function of CTC cell, and while, docks with micro-fluidic multicolor fluorescence CTC calculating instrument; Secondly, CTC is carried out to the peripheral blood of patients with colorectal cancer obtained and catch design test.
Wherein the detailed process of Magneto separate and dyeing is: extract 5.0mL peripheral blood sample, after 800 revs/min of centrifugings, extracts upper serum mixed liquor; Add ferrofluid (mass concentration 0.5%) and 2.0mL phosphate buffer (concentration 0.1mol/L) that 50 μ L mark anti-EpCAM antibody (Sigma Co., USA); Magnetic is hatched; Magnetic resolution extracts unlabeled cells; Again be suspended in phosphate buffer, add DAPI dyeing liquor (the green skies are biological), CK19 dyeing liquor (Ebioscience respectively, article No. 53-9898-82) and CD45 dyeing liquor (Ebioscience, article No. 12-0459-42) give dyeing 15min, magnetic resolution collects the cell after dyeing respectively.
Cell-anti-the CK-PE of mark fluorescent staining reagent identifies cell within a cell keratoprotein 8,18 and/or 19; Nuclear material dyes by DAPI; Anti-CD45-APC identifies white blood cell; Magneto separate washing is carried out in order to follow-up use to the CTC cell after dyeing.Wherein, fluorescent dye adopts three kinds of fluorescent reagents to be respectively DAPI dye core (blue light), and the anti-CD45 of PE-marks leucocyte (ruddiness), FITC-anti-CK protein labeling epithelial tumor cell (green glow).
The sample size controlling last micro-fluidic multicolor fluorescence CTC detector is 5 μ L, solid matter content≤0.5%.
(2) structure of micro-fluidic chip 3
Micro-fluidic chip 3 mainly utilizes micro-fluidic chip to carry out precise manipulation to the accurate manipulation ability of small items (cell) in micro-meter scale fluid to the CTC cell adding chip, by the Exact Design of micro-fluidic chip, realize CTC monolayer in chip, in conjunction with multicolor fluorescence analysis instrument, realize fluorescence analysis and the quick counter of individual cells.
Micro-fluidic chip 3 is by employing standard Soft lithograph technology (softlithography), and spin coating photoresist is in silicon chip surface, and solidification, develops after uv-exposure, makes micro-fluidic chip template.Silicone (PDMS) (traditional Chinese medicines chemical reagent company limited) is 5:1 mixing by A:B, and degassed in a vacuum, pours into and is equipped with in the double dish of mould, the thickness of falling glue 5 millimeters, be separated by PDMS after 80 DEG C of 60min that are heating and curing with mould.Punching, aims at upper and lower two-layer PDMS, bonding.80 DEG C are toasted 1 hour.Micro-fluidic chip 3 comprises the part composition such as pipeline 20 (comprising inwall space pattern to modify), injection port, outlet, and pipeline 20 inwall side or bilateral carry out finishing.Sample introduction, washing, shift and go out sample and all operated by the programmed digital code of miniature electromagnetic valve by control system, sample size is accurately controlled by miniature volume pump and built-in chip type valve system.
(3) structure of micro-fluidic multicolor fluorescence CTC detector
The fluorescence light source system of micro-fluidic multicolor fluorescence cell counter comprises micro-fluidic chip 3 (monolayer covering) and sampling device; Light source 1 device near micro-fluidic chip 3; The fluorescence detection device of a close described micro-fluidic chip 3 and described light source 1 device; A signal processor be connected with fluorescence detection device.Micro fluidic device wherein covers to carry out fluorescently-labeled circulating tumor cell, and therefore cell by the impinging light beam from described light source row, and can send fluorescence signal by process; These different fluorescence signals can turn back to processor and carry out Treatment Analysis, statistics.Micro-fluidic multicolor fluorescence CTC calculating instrument schematic diagram is shown in by its principle of device schematic diagram.
(4) application of micro-fluidic multicolor fluorescence CTC detector
Micro-fluidic fluorecyte calculating instrument is adopted to carry out CTC counting statistics and analysis the peripheral blood of colorectal cancer patients 10 example of originating above, result shows the circulating tumor cell number that 15 routine cancer patients come out and is respectively 6,16,92,213,112,66,65,26,17,9, the result of its result and artificial counting is without significant difference, error range within ± 5.0%, and conforms to the clinical data of reality.The consistance of above-mentioned micro-fluidic multicolor fluorescence cytometric analysis result and traditional pathology artificial counting result shows, calculating instrument of the present invention can realize the detection realizing circulating tumor cell in peripheral blood rapidly and accurately.The relation research of early diagnosis, metastases, chemotherapy effect, prognosis can be applied to; Also may be used for the detection of circulating tumor cell in postoperative patient blood, testing result and the relation predicting Preventive and prognosis thereof, the Primary Study of testing result and medication predicting relation; Or further according to just controlling and the detection analysis of postoperative patient, tentatively setting up early diagnosis, judging recurrence, direction of medication usage, the relevant criterion of prediction prognosis or guidance program.
Embodiment 2
The workflow of the fluorescence imaging number system of micro-fluidic multicolor fluorescence cell counter.
Tumour cell fluorescence detector key components, the precision architecture of fluoroscopic imaging systems is shown in accompanying drawing 1.The present embodiment adopts omnipotent infinity correcting optical system.Light is by becoming parallel beam after infinity chromatic aberration correction object lens 8, and this parallel beam is imaged onto CCD sensitive chip through pipe mirror 16.Can optical accessory be added and not affect total magnification between object lens 8 and pipe mirror 16.
This fluorescent microscope can distinguish cancer cell and normal cell fast, and job step is as follows:
1. tested cell is respectively with the dyeing of DAPI, FITC and PE fluorescent reagent;
2. tested loading cells is in micro-fluidic chip 3, is placed in by micro-fluidic chip 3 on objective table 4;
3. dichroic mirror 214 is shifted out light path, optical filter runner 15 is switched to neutral gear (not installing optical filter);
4. lighting source 1 and condenser 2 device are for the tested cell that throws light on, observable cellular morphology.Open lighting source 1, regulate the aperture of condenser 2, until brightness is moderate;
5. regulate coarse adjustment knob and the fine tuning knob of microscope stand, until image clearly can be photographed by video camera 17;
This step can be used for observation of cell form.
6. close lighting source 1.
7. optical filter runner 15 is switched to 461nm gear;
8. dichroic mirror 111 is installed in light path, and dichroic mirror 214 is moved out of light path;
9. open 405nm laser instrument A 9, laser is irradiated on tested cell through beam expanding lens A10,40 times of object lens 8, and produce blue-fluorescence after being excited by the cell of DAPI fluorochrome label, centre wavelength is 461nm.Now can be observed blue-fluorescence image by video camera 17;
The blue-fluorescence that this step produces for the cell observing DAPI fluorochrome label.
10. optical filter wheel is switched to 525nm gear;
11. dichroic mirrors 111 are moved out of light path, and dichroic mirror 214 is installed in light path;
12. open 488nm laser instrument B 12, and laser is irradiated on tested cell through beam expanding lens B13,40 times of object lens 8, and produce green fluorescence after being excited by the cell of FITC fluorochrome label, centre wavelength is 525nm.Now can be observed green fluorescence image by video camera (17);
The green fluorescence that this step produces for the cell observing FITC fluorochrome label.
10. optical filter runner 15 is switched to 670nm gear;
11. dichroic mirrors 111 are moved out of light path, and dichroic mirror 214 is installed in light path;
12. open 488nm laser instrument B12, and laser is irradiated on tested cell through beam expanding lens B13,40 times of object lens 8, and produce red fluorescence after being excited by the cell of PE fluorochrome label, centre wavelength is 670nm.Now can be observed red fluorescence images by video camera 17;
The red fluorescence that this step produces for the cell observing PE fluorochrome label.
Claims (10)
1. a micro-fluidic multicolor fluorescence cell counter, comprises micro-fluidic chip system, fluoroscopic imaging systems and analysis meter number system; The controller (18) that described micro-fluidic chip system comprises micro-fluidic chip (3), computing machine (19) controls and pipeline (20); Described fluorescence light source system comprises two kinds or more laser instruments, as the object lens (8) of laser instrument A (9) and laser instrument B (12), fluorescence detection and the video camera (17) of signal transacting; Described analysis meter number system comprises computing machine (19).
2. cell counter according to claim 1, is characterized in that, described micro-fluidic chip system is also alternatively containing objective table (4), motor (5), one of guide rail (6) and screw mandrel (7) or more.
3. cell counter according to claim 1, it is characterized in that, described fluoroscopic imaging systems is also alternatively containing one of lighting source (1), condenser (2), object lens (8), beam expanding lens A (10), dichroic mirror 1 (11), beam expanding lens B (13), dichroic mirror 2 (14), optical filter runner (15) or more.
4. cell counter according to claim 1, is characterized in that, described object lens (8) preferably 40 times of Liars, and described laser instrument A (9) is 405nm, described laser instrument B (12) preferably 488nm preferably.
5. cell counter according to claim 1, is characterized in that, the material of described micro-fluidic chip (3) is silicone; There is three-dimensional structure; Chip thickness is less than 50 microns; In chip, sample capacity is less than 10 microlitres; Chip surface adopts EpCAM antibody modification.
6. cell counter according to claim 1, is characterized in that, described fluorescence light source system establishes two kinds of fluorescence excitation light sources; The cell that described laser instrument (A) fluorescent exciting source excitation is dyeed by 4', 6-diamidino-2-phenylindone (DAPI), produces blue-fluorescence; The cell that described laser instrument (B) fluorescent exciting source excitation is dyeed by fluorescein isothiocynate (FITC) and phycoerythrin (PE), produces green fluorescence and ruddiness fluorescence respectively.
7. a preparation method for micro-fluidic multicolor fluorescence cell counter, comprises and prepares micro-fluidic chip (3) with following steps,
1) spin coating photoresist is in silicon chip surface, and solidification, develops after uv-exposure, makes micro-fluidic chip (3) template;
2) silicone (PDMS) is 5:1 mixing by A:B, degassed in a vacuum, pours into and is equipped with in the double dish of mould, the thickness of falling glue 5 millimeters, be separated by PDMS after 80 DEG C of 60min that are heating and curing with mould;
3) punch, upper and lower two-layer PDMS is aimed at, bonding; 80 DEG C are toasted 1 hour.
8. cell counter according to claim 1 is used for counting and the graphical analysis of circulating tumor cell in blood.
9. cell counter according to claim 1 is used for colorectal cancer circulating tumor cell counting.
10. the application of cell counter according to claim 1, comprises and carries out circulating tumor cell Magneto separate and qualification in advance; Or, be included in micro-fluidic chip (3) and carry out cellular morphology and fluorescence labeling state analysis.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106732839A (en) * | 2016-12-30 | 2017-05-31 | 天津禄浩科技股份有限公司 | A kind of cellular fat particle detections chip and its detection reagent |
| CN109762868A (en) * | 2019-01-15 | 2019-05-17 | 大连大学 | A method of based on the micro-fluidic chip Accurate Analysis cell cycle |
| CN109870401A (en) * | 2019-02-28 | 2019-06-11 | 西安理工大学 | A kind of acquisition method of flow cytometer and super-resolution cell image |
| CN109991205A (en) * | 2019-05-05 | 2019-07-09 | 中国科学院重庆绿色智能技术研究院 | A kind of counting algorithm of circulating tumor cell and application |
| CN110199186A (en) * | 2016-11-22 | 2019-09-03 | 芯易诊有限公司 | Whole blood count measurement method |
| CN111398237A (en) * | 2020-05-07 | 2020-07-10 | 成都量蚁科技有限公司 | Cell detection device, application and cell classification detection method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120129192A1 (en) * | 2010-11-22 | 2012-05-24 | Nauganeedles Llc | Apparatus and Methods for Detection of Tumor Cells in Blood |
| CN102911860A (en) * | 2012-09-29 | 2013-02-06 | 武汉斯坦姆赛尔生物技术有限公司 | Portable fluorecyte counting device |
| CN103439242A (en) * | 2013-09-06 | 2013-12-11 | 华南师范大学 | Microfluidic system and method for detecting and screening single beam biological cells |
| CN204269525U (en) * | 2014-09-01 | 2015-04-15 | 上海柏慧康生物科技有限公司 | A kind of micro-fluidic multicolor fluorescence cell counter |
-
2014
- 2014-09-01 CN CN201410441265.5A patent/CN104280329A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120129192A1 (en) * | 2010-11-22 | 2012-05-24 | Nauganeedles Llc | Apparatus and Methods for Detection of Tumor Cells in Blood |
| CN102911860A (en) * | 2012-09-29 | 2013-02-06 | 武汉斯坦姆赛尔生物技术有限公司 | Portable fluorecyte counting device |
| CN103439242A (en) * | 2013-09-06 | 2013-12-11 | 华南师范大学 | Microfluidic system and method for detecting and screening single beam biological cells |
| CN204269525U (en) * | 2014-09-01 | 2015-04-15 | 上海柏慧康生物科技有限公司 | A kind of micro-fluidic multicolor fluorescence cell counter |
Non-Patent Citations (5)
| Title |
|---|
| LEOPOLD G. KOSS等: "《KOSS诊断细胞学及其组织病理学基础》", 30 June 2009, 西安: 世界图书出版西安公司 * |
| 张金玲: "基于微流控芯片的免疫反应快速检测系统研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
| 徐永前等: "《恶性积液诊断与治疗》", 30 September 2000, 北京：中国科学技术出版社 * |
| 李丽秀: "共聚焦激光诱导荧光微流控芯片检测系统的研究", 《豆丁网》 * |
| 王晓波等: "《复方黄黛制剂 临床与作用机理》", 31 January 2011, 北京：北京科学技术出版社 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110199186A (en) * | 2016-11-22 | 2019-09-03 | 芯易诊有限公司 | Whole blood count measurement method |
| CN106732839A (en) * | 2016-12-30 | 2017-05-31 | 天津禄浩科技股份有限公司 | A kind of cellular fat particle detections chip and its detection reagent |
| CN106732839B (en) * | 2016-12-30 | 2022-04-26 | 天津禄浩科技股份有限公司 | Cell fat particle detection chip and detection reagent thereof |
| CN109762868A (en) * | 2019-01-15 | 2019-05-17 | 大连大学 | A method of based on the micro-fluidic chip Accurate Analysis cell cycle |
| CN109870401A (en) * | 2019-02-28 | 2019-06-11 | 西安理工大学 | A kind of acquisition method of flow cytometer and super-resolution cell image |
| CN109870401B (en) * | 2019-02-28 | 2021-09-10 | 西安理工大学 | Flow cytometer and super-resolution cell image acquisition method |
| CN109991205A (en) * | 2019-05-05 | 2019-07-09 | 中国科学院重庆绿色智能技术研究院 | A kind of counting algorithm of circulating tumor cell and application |
| CN111398237A (en) * | 2020-05-07 | 2020-07-10 | 成都量蚁科技有限公司 | Cell detection device, application and cell classification detection method |
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