CN1161613C - Immune cell microfluid array - Google Patents
Immune cell microfluid array Download PDFInfo
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- CN1161613C CN1161613C CNB021384746A CN02138474A CN1161613C CN 1161613 C CN1161613 C CN 1161613C CN B021384746 A CNB021384746 A CN B021384746A CN 02138474 A CN02138474 A CN 02138474A CN 1161613 C CN1161613 C CN 1161613C
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Abstract
The present invention relates to an immune cell microfluid array which belongs to the technical field of cell immune inspection. A bottom board of a macromolecular compound of the immune cell microfluid array is provided with micro array counterbores which are orderly arranged; the counterbores are communicated through micro pipelines; a micro pipeline between two adjacent counterbores is arranged slantways; a nozzle on the counterbore with a front sequence is higher than the nozzle on the adjacent counterbore with a back sequence. By using the present invention, a cell with corresponding leukocyte differentiation antigens can be inspected by one-step operation. Consequently, the kinds of the leukocyte differentiation antigens of the surface of the cell, an eat and flow rule of the leukocyte differentiation antigens in the process of cell differentiation, etc. can be judged. The present invention greatly improves work efficiency. Moreover, so long as a culture agent is added, the growth state of the cell with the different leucocyte differentiation antigens can be observed through the culture of certain time. Consequently, the present invention can integrally appraise whether the cell/ an organism is normal or not, which is favorable for integrally appraising the integral health levels of immune functions, inflammation generation, tumor cell metastasis, etc. of an organism.
Description
Technical field
The present invention relates to a kind of verifying attachment of immunocyte, especially the microfluidic arrays of immunocyte belongs to cellular immunity inspection technology field.
Background technology
Along with the development and the various The application of new technique of a series of technology such as modern biology, biological chemistry and immunology, the research of human leukocytes differentiation antigen has obtained remarkable progress.
Studies show that leukocyte differentiation antigen not only at leucocyte (as lymphocyte, monocyte, granulocyte etc.) surface expression, also is distributed on the cell membrane of thymocyte, myeloid cell, stem cell, epithelial cell etc.It is the ripe cell surface marker for different pedigrees and different phase of cell normal differentiation, and occurs or disappearance with the cell activation degree.They mostly are protein or the glycoprotein of wearing film, contain after birth outskirt, transmembrane domains and cytoplasmic domain; Some leukocyte differentiation antigen be with phosphatidylinositols connected mode " anchor " on cell membrane; The minority leukocyte differentiation antigen is the carbohydrates haptens.According to distributing and function existing 9 big class, that is: the 1.T cells that substantially it are divided into; 2.B cell; 3. myeloid cell and monocyte; 4. blood platelet; 5.NK cell and non-pedigree cell; 6. active antigen; 7. vascular endothelial cell; 8. adhesion factor; 9. cytokine receptor.In organism physiology and pathologic process, leukocyte differentiation antigen not only participates in discerning antigen, capture antigen, promote the interaction between immunocyte and antigen or immune molecule, and the effect of sticking between the mediation immunocyte, between immunocyte and matrix, the identification of immune response, activation and effective stage all play a significant role, in the regulation and control of hematopoietic cell differentiation and hematopoiesis, inflammation takes place and the aspects such as transfer of tumour cell also play an important role simultaneously.Therefore, the research of leukocyte differentiation antigen is not only some leukocytic research, also can be reflected the general level of the health of whole machine body simultaneously, significant.
At present, the detection of different leukocyte differentiation antigens is adopted usually cell sample to be checked is fixed on the detection base plate, drips the method for certain antibody and indicant again.When antibody combines with corresponding antigen on the cell membrane and produces specific reaction, can detect and contain certain antigen in the sample cell.The subject matter one that this prior art exists is that checkability is low, because fixed leukocyte differentiation antigen has 166 kinds of molecular groups and numerous subgroup, check is very loaded down with trivial details one by one; The 2nd, the leucocyte after fixing loses activity, and can not cultivate, and therefore can't further observe the growth conditions of the cell with different leukocyte differentiation antigens.
Summary of the invention
The objective of the invention is to: propose a kind of immunocyte microfluidic arrays that can greatly improve the leukocyte differentiation antigen checkability.
In order to reach above purpose, the basic technical scheme of immunocyte microfluidic arrays of the present invention is: comprise the macromolecular compound base plate, be shaped on the microarray counterbore of arranging in order on this base plate, be communicated with by microchannel between each counterbore, microchannel between the two adjacent counterbores is obliquely installed, ordering is higher than ordering at the back mouth of pipe on counterbore at the mouth of pipe on the preceding counterbore, and described microarray counterbore bottom is fixed the antibody of anti-leukocyte differentiation antigen respectively in an orderly manner.
When the leukocyte differentiation antigen that carries out cell is analyzed, fix the antibody of various anti-leukocyte differentiation antigens earlier respectively in an orderly manner in each counterbore bottom of microarray base plate, then test sample (cell) is transported to the microarray counterbore the preceding that sorts continuously, each microarray counterbore that cell liquid will have been fixed different antibodies by flowing through one by one behind the forward direction according to the order of sequence is until flowing out from the counterbore of position, end.When being detained, just can concluding and contain corresponding differentiation antigen in the sample cell when the antibody generation specificity association reaction in sample cell and certain counterbore.Like this, single job just can detect the leukocyte differentiation antigen type that the sample cell is had, thereby judges the kind of cell surface leukocyte differentiation antigen, the leukocyte differentiation antigen growth and decline rule in the cell differentiation procedure etc., and work efficiency improves greatly.And because sessile antibody in the counterbore, when its combine with corresponding antigen on the cell membrane with cell separation, fixing after, can keep leukocytic activity, as long as add nutrient chemical, cultivation through certain hour, just can observe the growth conditions of cell, thereby whether pair cell/body normally carries out the overall evaluation, be beneficial to the holistic health levels such as immunity function, inflammation generation and tumour cell transfer of comprehensive evaluation body with different leukocyte differentiation antigens.
On above basic technical scheme basis, the further improvement of the present invention is that the microarray counterbore two ends on the macromolecular compound base plate are shaped on the first reservoir and last position reservoir that is communicated with the first counterbore and position, end counterbore respectively, connects by peristaltic pump between two reservoirs.Like this, after the cell liquid/nutrient solution that flows out from end position counterbore enters position, end reservoir, can be transported to again and the first reservoir by peristaltic pump, each microarray counterbore of flowing through one by one and having fixed different antibodies again from the first counterbore, until from the counterbore of end position, flowing out, each microarray counterbore of flowing through one by one, formation circulates, make the amount of liquid unanimity in every each counterbore on the mocroarray plate, more help obtaining desirable assay.
Description of drawings
The present invention is further illustrated below in conjunction with accompanying drawing.
Fig. 1 is the structural representation of one embodiment of the invention.
Embodiment
Embodiment one
The immunocyte microfluidic arrays of present embodiment is shaped on immunity (cell) the microarray counterbore 4 of 15 * 20 (or 10 * 20) as shown in Figure 1 on the macromolecular compound base plate 1 of acellular poison.The thickness of base plate 1 is 7～10mm, and the aperture of each counterbore 4 is 2mm ± 0.2mm, and the spacing of each adjacent counterbore is 2～3mm, and the microchannel 2 by diameter 0.1～0.5mm communicates each other.The connection microchannel of two adjacent counterbores is tilting according to following rule, and the microchannel opening of ordering on preceding hole be apart from hole bottom 0.8～1.0mm, ordering on metapore the microchannel opening apart from bottom, hole 0.1～0.25mm; By that analogy, continue, form reciprocal S shape circulatory flow, all counterbores are linked to each other.Experiment shows that above design parameter is suitable for the cellular immunity check.
At the two ends of microarray base plate 1, the foreign side of promptly the first counterbore and position, end counterbore respectively is shaped on a reservoir 5 that can hold 2ml liquid.Wherein, the reservoir that links to each other with the first counterbore of microarray claims A groove, and the reservoir that links to each other with position, end counterbore claims B groove, connects with peristaltic pump 3 between two grooves.Like this, liquid can be from B groove pump to the A groove, and the liquid of A groove is then gone into the first counterbore, 2 holes of microarray by the run by gravity of hydraulic pressure ... until position, end counterbore, be back to the B groove, form circulation, make the amount of liquid unanimity in each counterbore on the mocroarray plate.
When the leukocyte differentiation antigen that carries out cell is analyzed, fix the antibody of various anti-leukocyte differentiation antigens earlier respectively in an orderly manner in each counterbore bottom of mocroarray plate, then test sample (cell) is added in the A groove and the nutrient chemical mixing, through the channel flow that communicates through each counterbore.According to the difference of the leukocyte differentiation antigen of expressing on the cell membrane, test sample (cell) combines with specific antibody in the corresponding counterbore and is fixed.According to research purpose, can directly examine under a microscope, analyze the leukocyte differentiation antigen that test sample (cell) is had.
The immunocyte microfluidic arrays of present embodiment is applicable to the kind of the leukocyte differentiation antigen of separation, fixing and final identification of cell surface expression, in the hope of analysis of cells activation degree, leukocyte differentiation antigen growth and decline rule, the differential growth situation of different cells is with the holistic health level of comprehensive evaluation body.
Embodiment two
Present embodiment is identical with the basic condition of embodiment one, and difference is that therefore whole process is operated in gnotobasis because need pair cell to carry out further researchs such as growth and breeding.For the cultivation that guarantees the later stage reaches aseptic effect, the macromolecular compound base plate is furnished with the lid of isolation.Finish with embodiment one identical operations after, add a cover and put into incubator, and the regular recycling replacing nutrient chemical by A groove → B groove, incubation time is determined as required.Can obtain further related check result like this.
In addition to the implementation, the present invention can also have other embodiments.All employings are equal to the technical scheme of replacement or equivalent transformation formation, all drop in the protection domain of requirement of the present invention.
Claims (6)
1. immunocyte microfluidic arrays, comprise the macromolecular compound base plate, it is characterized in that: be shaped on the microarray counterbore of arranging in order on the described base plate, be communicated with by microchannel between each counterbore, microchannel between the two adjacent counterbores is obliquely installed, ordering is higher than ordering at the back mouth of pipe on counterbore at the mouth of pipe on the preceding counterbore, and described microarray counterbore bottom is fixed the antibody of anti-leukocyte differentiation antigen respectively in an orderly manner.
2. according to the described immunocyte microfluidic arrays of claim 1, it is characterized in that: the microarray counterbore two ends on the described macromolecular compound base plate are shaped on the first reservoir and position, the end reservoir that is communicated with the first counterbore and position, end counterbore respectively, connect by peristaltic pump between two reservoirs.
3. according to claim 1 or 2 described immunocyte microfluidic arrays, it is characterized in that: be communicated with the reciprocal S shape circulatory flow of formation by microchannel between the described microarray counterbore.
4. according to claim 1 or 2 described immunocyte microfluidic arrays, it is characterized in that: the aperture of described microarray counterbore is 2mm ± 0.2mm, and the spacing of each adjacent counterbore is 2～3mm.
5. according to claim 1 or 2 described immunocyte microfluidic arrays, it is characterized in that: the diameter of described microchannel is 0.1～0.5mm.
6. according to claim 1 or 2 described immunocyte microfluidic arrays, it is characterized in that: described macromolecular compound base plate is furnished with the lid of isolation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021384746A CN1161613C (en) | 2002-10-22 | 2002-10-22 | Immune cell microfluid array |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021384746A CN1161613C (en) | 2002-10-22 | 2002-10-22 | Immune cell microfluid array |
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| CN1403817A CN1403817A (en) | 2003-03-19 |
| CN1161613C true CN1161613C (en) | 2004-08-11 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNB021384746A Expired - Fee Related CN1161613C (en) | 2002-10-22 | 2002-10-22 | Immune cell microfluid array |
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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9121843B2 (en) | 2007-05-08 | 2015-09-01 | Trustees Of Boston University | Chemical functionalization of solid-state nanopores and nanopore arrays and applications thereof |
| EP2483680A4 (en) | 2009-09-30 | 2014-01-01 | Quantapore Inc | Ultrafast sequencing of biological polymers using a labeled nanopore |
| CN102952752B (en) * | 2011-08-29 | 2014-07-09 | 东南大学 | Culture inspection device for cancer cells |
| US9651539B2 (en) | 2012-10-28 | 2017-05-16 | Quantapore, Inc. | Reducing background fluorescence in MEMS materials by low energy ion beam treatment |
| WO2014190322A2 (en) | 2013-05-24 | 2014-11-27 | Quantapore, Inc. | Nanopore-based nucleic acid analysis with mixed fret detection |
| JP6740222B2 (en) | 2014-10-10 | 2020-08-12 | クアンタポール， インコーポレイテッド | Nanopore-based polymer analysis with fluorescent labels quenching each other |
| EP3209800B1 (en) | 2014-10-24 | 2022-02-16 | Quantapore, Inc. | Efficient optical analysis of polymers using arrays of nanostructures |
| EP3482196B1 (en) | 2016-07-05 | 2022-02-23 | Quantapore, Inc. | Optically based nanopore sequencing |
| CN105929152B (en) * | 2016-07-15 | 2018-06-26 | 安徽国科生物科技有限公司 | A kind of intelligent detecting instrument for Procalcitonin detection |
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2002
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