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Single cell array chip and screening method for ion channel drugs

An array chip, ion channel technology, applied in the field of ion channel drug screening, can solve the problems of high technical threshold, inaccuracy of FlexStation3, poor reproducibility of results, etc., to reduce false positive rate/false negative rate and improve results. Reproducibility, the effect of reducing sample consumption

Active Publication Date: 2020-04-03
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Traditional ion channel drug screening platforms include traditional calcium fluorescence high-throughput screening technology (FlexStation 3) and whole-cell patch clamp technology. Problems such as expensive equipment
The traditional calcium fluorescence high-throughput screening technology measures the total fluorescence value of all cells in a single well of a well plate (such as a 96-well plate). First, the number of cells is uncertain. In addition, the adhesion effect of the cells is not good. Repeated liquid addition steps in the microplate reader may cause cells to fall off the bottom of the plate. These uncertain factors also cause inaccurate and difficult to repeat results of FlexStation 3
Active compounds screened by the traditional calcium fluorescence high-throughput method are usually difficult to be effectively verified on the gold index patch clamp, indicating that the accuracy of the screening results using the traditional calcium fluorescence technology is not high, and there are a large number of false positive or false negative results.
The whole-cell patch clamp technique is not suitable for large-scale screening due to its high technical threshold and low throughput.

Method used

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  • Single cell array chip and screening method for ion channel drugs
  • Single cell array chip and screening method for ion channel drugs
  • Single cell array chip and screening method for ion channel drugs

Examples

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preparation example Construction

[0059] According to the preparation method of the present invention, in step (1), the design drawings of the lower microporous chip and the upper microchannel chip can be drawn by using AutoCAD software. Preferably, the resolution of the high-resolution transparent mask can be 25000dpi or above, so as to ensure the fidelity of the graphics of the present invention.

[0060] According to the preparation method of the present invention, in step (2), the silicon wafer is rinsed with absolute ethanol, boiled with Piranha solution, rinsed with ultrapure water to neutrality, dried and other pretreatments, and then applied to subsequent coating deal with. Described silicon wafer pretreatment method can be: first silicon wafer is rinsed with absolute ethanol, to remove the impurity adhering to silicon wafer surface, blow dry; Then silicon wafer is placed in Piranha solution (concentrated sulfuric acid:hydrogen peroxide=3 : 1) Heating and boiling, the heating time is 10-50min, prefera...

Embodiment 1

[0091] Example 1 - Preparation of single cell array chip

[0092] The structure of the single-cell array chip: the upper surface of the lower microwell chip is provided with a number of cylindrical micro-pits, the diameter of the micro-pit is 25 μm, the depth of the micro-pit is 40 μm, and the distance between the centers of adjacent micro-pit is 80 μm, the micro-pit An array of micro-pits arranged in a linear equidistant manner is formed. 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...

Embodiment 2

[0102] 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 and a screening method for ion channel drugs. The single cell array chip includes a lower layer micro-well chip and an upper layer micro-channel chip,and the lower layer micro-well chip and the upper layer micro-channel chip are bonded together; the upper surface of the lower layer micro-well chip is provided with micro-pits, and the center distances of the adjacent micro-pits are 30-90 [mu]m; and the material of the single cell array chip is selected one or more of high-molecular polymer, silicon, glass, quartz and paper. The method for screening ion channel drugs by utilizing the single cell array chip can obtain higher single cell capture rates, and the heterogeneity of single cells can be fully considered; and the screening method can enhance the accuracy and reproducibility of single cell level drug screening results, significantly reduce false positive rates / false negative rates and increase the screening efficiency of the ion channel drugs.

Description

technical field [0001] The invention relates to a single cell array chip, and also relates to a screening method for ion channel drugs. 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 screening platforms include traditional calcium fluorescence high-throughput screening technology (FlexStation 3) and whole-cell patch clamp technology. Problems such as expensive equipment. The traditional calcium fluorescence high-throughput screening technology measures the total fluorescence value of all cells in a single well of a well plate (such as a 96-well plate). First, the number of cells is uncertain. In ad...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12Q1/02
CPCC12M23/16G01N33/5008G01N2500/10
Inventor 艾晓妮姜勇卢文博屠鹏飞
Owner PEKING UNIV
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