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Single-cell array chip and preparation method and application thereof

An array chip and single-cell technology, which is applied in the field of preparation of the single-cell array chip, can solve problems such as high-throughput screening of unseen ion channel drugs, reduce sample consumption, improve result accuracy, and reduce false positive rates / False Negative Rate Effect

Active Publication Date: 2020-04-17
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods are mostly used in genome amplification, single cell in situ culture, proliferation, induced differentiation, content extraction, etc., and there are no reports on high-throughput screening of ion channel drugs

Method used

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  • Single-cell array chip and preparation method and application thereof
  • Single-cell array chip and preparation method and application thereof
  • Single-cell array chip and preparation method and application thereof

Examples

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preparation example Construction

[0050]

[0051] The single cell array chip of the present invention can be prepared by various methods.

[0052] According to one embodiment of the present invention, the single-cell array chip is prepared by a soft lithography PDMS injection molding method, and the method includes the following steps:

[0053] (1) Draw the design diagrams of the lower microwell chip and the upper microchannel chip respectively, and make a high-resolution transparent mask with micropit array graphics and a high-resolution transparent mask with microfluidic channel graphics respectively;

[0054] (2) Coat the negative photoresist SU8 2007 on the silicon wafer with a coating thickness of 5-10 μm and cure to obtain an adhesion layer; then coat SU8 2050 on the surface of the adhesion layer and coat The thickness of the cloth is 20-60 μm, and it is pre-baked to obtain photoresist;

[0055] (3) Place the pattern of the high-resolution transparent mask with the micropit array pattern on the photor...

Embodiment 1

[0073] The single-cell array chip is formed by bonding a lower microwell chip and an upper microchannel chip. The upper surface of the lower microporous chip is provided with a number of cylindrical micro-pits, the diameter of the micro-pits is 25 μm, the depth of the micro-pits is 40 μm, and the distance between the centers of adjacent micro-pits is 80 μm, and the micro-pits form a straight line equidistant Uniform array of micropits. The upper microchannel chip is provided with two microfluidic channels, and the two ends of each microfluidic channel are respectively provided with inflow through holes and outflow through holes. The microfluidic channel is located on the lower surface of the upper microchannel chip and communicates with the micropit; the inflow through hole and the outflow throughhole communicate the microfluidic channel with the upper surface of the upper microchannel chip. The cross-section of the microfluidic channel is rectangular, the length of the micro...

Embodiment 2

[0075] Except that the diameter of the micropit is 28 μm, other parameters are the same as in Example 1.

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Abstract

The invention discloses a single-cell array chip as well as a preparation method and an application thereof. The single-cell array chip comprises a lower-layer micropore chip and an upper-layer microchannel chip, wherein the lower-layer micropore chip and the upper-layer microchannel chip are bonded together; the upper surface of the lower microporous chip is provided with a plurality of micro-pits, and the distance between the centers of adjacent micro-pits is 30-90 [mu]m. The single-cell array chip provided by the invention can obtain higher single-cell capture rate and higher single-cell level drug screening efficiency.

Description

technical field [0001] The invention relates to a single-cell array chip, and also relates to a preparation method and application of the single-cell array chip. Background technique [0002] Ion channels are a class of transmembrane biomacromolecules with selective permeability to ions, which regulate various physiological activities of the body. Ion channel dysfunction can induce a variety of "ion channelopathies", such as epilepsy, arrhythmia, diabetes and so on. Drug development targeting ion channels is a hot topic in academia and industry. [0003] Traditional ion channel drug target screening platforms include traditional calcium fluorescence high-throughput screening technology (FlexStation 3) and whole-cell patch clamp technology, but both of these technologies have high reagent consumption, poor reproducibility of results, and high technical threshold , expensive equipment and other issues. Active compounds screened by the traditional calcium fluorescence high-t...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12Q1/02B01L3/00
CPCC12M23/16C12M23/12G01N33/5008B01L3/502707B01L3/502761B01L2200/0652B01L2300/0887
Inventor 艾晓妮姜勇卢文博王克威屠鹏飞吴洋张新然
Owner PEKING UNIV
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