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Cell-microbead pairing capture method applied to unicellular omics analysis

A single-cell, cell-based technology, applied in the direction of sterilization methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems of cross-contamination of capture structures, large sample consumption, complex structures, etc., and achieve low cost, Ease of operation and high capture efficiency

Pending Publication Date: 2022-08-09
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a cell-microbead paired capture method applied to single-cell omics analysis, so as to solve the problems of cross-contamination, large sample consumption, complex structure, and common problems in the existing single-cell omics analysis platform. low volume problem

Method used

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  • Cell-microbead pairing capture method applied to unicellular omics analysis
  • Cell-microbead pairing capture method applied to unicellular omics analysis
  • Cell-microbead pairing capture method applied to unicellular omics analysis

Examples

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Embodiment 1

[0037] Example 1 A cell-microbead paired capture method

[0038] combine Figure 1 to Figure 3 As shown, a cell-microbead paired capture method provided according to a preferred embodiment of the present invention is shown, comprising the following steps:

[0039] S1: First provide a figure 1 The microfluidic capture device shown, the microfluidic capture device is composed of a particle capture layer and a substrate layer, and the two layers can be separated; the particle capture layer is provided with an inlet, an outlet, and one-to-one paired capture cells - Several capture microstructures of the microbeads distributed in an array.

[0040] S2: Pass the cell suspension and microbead suspension into the particle capture layer sequentially from the inlet, and realize the capture of a single cell and a single microbead in sequence in the capture microstructure, thereby forming a large-scale cell-bead paired array .

[0041] S3: After the paired capture is completed, contro...

Embodiment 2

[0051] Embodiment 2 A microfluidic capture device and cell-microbead paired capture method

[0052] combine Figure 4 to Figure 5 As shown, a microfluidic capture device provided according to this preferred embodiment is shown, and the microfluidic capture device can capture cells-microbeads by a hydrodynamic method, and the detailed description is as follows:

[0053] The microfluidic capture device is composed of a particle capture layer and a substrate layer, and the two layers can be separated. like Figure 4 As shown, the particle capture layer sequentially includes from upstream to downstream: an inflow zone 1 , a capture zone 2 , and an outflow zone 3 .

[0054] The inflow area 1 includes: an inlet 4 , a branch channel 5 , and a pre-filter 6 . The inlet 4 is connected with several runners 5 of equal width. The runner 5 is symmetrical with the third channel 7 as the axis of symmetry. At the same time, a section is reserved at the rear end of the inflow area 1 to form ...

Embodiment 3

[0067] Example 3 Preparation of a Microfluidic Capture Device

[0068] According to this preferred embodiment, the present invention also provides a preparation method of a microfluidic capture device, the processing flow is as follows Figure 9 shown, the detailed description is as follows:

[0069] (1) Design and production of photolithography masks: draw the design diagram of the microfluidic device, and use high-precision printing to output multiple masks. A pattern is formed in the exposure step.

[0070] (2) Photolithography to prepare mold

[0071] Photolithography is micro-manufacturing with photoresist, mask and ultraviolet light, and the mold processing can be repeated and used. The specific process is as follows:

[0072] ①Clean the silicon wafer

[0073] ②Soft bake

[0074] ③Exposure: place the mask on the top of the silicon wafer, irradiate the silicon wafer with the photoresist spin-coated with ultraviolet light, the photoresist undergoes a photochemical react...

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Abstract

The invention provides a cell-microbead pairing capture method applied to unicellular omics analysis, and the method comprises the following steps: S1, providing a microfluidic capture device which comprises a separable particle capture layer and a substrate layer, the particle capture layer is internally provided with a plurality of capture microstructures for one-to-one pairing capture of cells-microbeads; s2, introducing a cell or microbead suspension from an inlet, and gradually completing one-to-one pairing capture of a single cell and a single microbead; s3, continuously filling the controllable curing solution from the inlet, and curing the controllable curing solution according to the characteristics of the solution, so that each cell-microbead capturing unit forms mutually isolated independent cavities; and S4, separating the substrate layer from the particle capture layer to form an open cell-microbead capture unit array, and carrying out single-cell omics analysis in the next step. According to the invention, a cell-microbead pairing capture method with high throughput, high capture efficiency, low cost and low pollution is provided for unicellular omics analysis.

Description

technical field [0001] The invention relates to the field of single-cell omics analysis, and more particularly to a cell-microbead paired capture method applied to single-cell omics analysis. Background technique [0002] Cells are the basic structural and functional units of organisms, and multicellular organisms are composed of various cells with different morphological functions. Cellular heterogeneity is a ubiquitous biological phenomenon, and even cells that appear to be identical may exhibit significant heterogeneity. Therefore, it is necessary to study the heterogeneity of cells at the single-cell level. It is necessary to establish a research platform to effectively analyze the chemical information of single cells, or to measure the response of single cells to external stimuli, so that people can fully understand the cell population. It can identify a small number of abnormal cells in a large number of cell populations, and provide a reliable scientific basis for th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12M1/12B01L3/00C12Q1/24
CPCC12M47/04C12M23/16C12M25/14C12M33/14B01L3/502707B01L3/502761C12Q1/24
Inventor 罗源邢晓星赵建龙王宁高文冰
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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