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High-throughput single cell capture and arrangement chip and method

A single-cell, high-throughput technology, applied in cell dissociation methods, biochemical equipment and methods, animal cells, etc., can solve problems such as low capture rate, inability to study cell interactions, etc., achieve high capture rate, price Inexpensive and simple method

Active Publication Date: 2018-06-15
BEIHANG UNIV
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  • Summary
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0020] In summary, the existing methods have the disadvantages of low capture rate or insufficient for studying cell-cell interactions

Method used

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  • High-throughput single cell capture and arrangement chip and method
  • High-throughput single cell capture and arrangement chip and method
  • High-throughput single cell capture and arrangement chip and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0062] Example 2: Comparison of single cell capture of myoblasts with prior art.

[0063] (1) Design a square porous membrane capture hole 1 ( figure 1 ) side length 25μm, square capture hole 2 ( figure 1 ) with a side length of 3 μm.

[0064] (2) Substrate production: After mixing the PDMS (Sylgard 184) prepolymer and the cross-linking agent at a mass ratio of 10:1, evenly throw a layer of 15 μm thick PDMS on the biological slide with a homogenizer at 4000 rpm Film 3 ( figure 2 ), cured at 65°C for 4 hours. The slides are biological slides 4 ( figure 2 ), the diameter of the glass climbing sheet can be 8-24mm, and the thickness is 0.17mm. The present embodiment selects the climbing sheet with a diameter of 24mm.

[0065] (3) Production of porous membrane: porous membrane 5 ( figure 2 ) contains cell capture well 1 and spreading well 2 ( figure 1 ), the captured cells can be different types of human or animal cells, and the pore size and distance of the porous membra...

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Abstract

The invention relates to a high-throughput single cell capture and arrangement chip and method and belongs to the field of materials and tissue engineering and particularly relates to high-throughputcapture and arrangement of single cells. The method comprises the following steps: making a substrate surface micro-pattern under the action of protein penetration, performing the high-throughput capture and arrangement of the single cells under the action of membrane pore limitation, and arranging and paving the cells on the complex micro-pattern. The high-throughput single cell capture and arrangement chip and method have the benefits that the capture rate of 85 percent or above of single cells can be realized, and the orderly arrangement and the orderly pavement of the cells on the specified protein pattern are realized; the influence caused by cellular heterogeneity during the large cell population study is avoided, and the method is simple and high in single cell capture rate, has a suitable follow-up culture environment, can be determined to be applied to the study on cell junction, intercellular interaction, cell signal transduction and the like at a single cell level and has avery important effect in the cell engineering and tissue engineering study.

Description

technical field [0001] The invention relates to a preparation method of a high-throughput single cell capture and arrangement chip and a capture and arrangement method, which belong to the field of materials and tissue engineering, and relate to the research on the capture and arrangement of single cells by combining the membrane pore method and the micropattern method, which can solve the problem of Due to the impact of cellular heterogeneity among large cell populations, subsequent single-cell analysis plays a pivotal role in the fields of genomics, transcriptomics, proteomics, and metabolomics. Background technique [0002] In traditional research, cell samples are often regarded as stable and uniform, ignoring the influence of intercellular heterogeneity. Experiments show that the average response of cell samples cannot accurately reflect the exact state of individual cells. So the study of single cells is very important. The capture of single cells is the premise of c...

Claims

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Application Information

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IPC IPC(8): C12M1/12C12M1/00C12N5/077
CPCC12M47/04C12N5/0658C12N2509/00C12N2533/52C12N2533/54
Inventor 孙艳浑婷婷樊瑜波王玮刘姚萍
Owner BEIHANG UNIV
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