Micro-fluidic chip for capturing single cells

A microfluidic chip and single-cell technology, applied in the field of microfluidics, can solve the problem of not achieving single-cell level capture, and achieve the effect of simple structure, flexible combination, and good function realization.

Active Publication Date: 2019-07-12
NORTHEASTERN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this device does not achieve single-cell capture,

Method used

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  • Micro-fluidic chip for capturing single cells
  • Micro-fluidic chip for capturing single cells
  • Micro-fluidic chip for capturing single cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1～3

[0074] In the following examples 1-3:

[0075] 1. The preparation process of the single cell capture microfluidic chip is as follows:

[0076] (1) Silicon wafer cleaning: standard silicon wafer cleaning, placed on a 200°C electric heating plate for 15 minutes, and dried;

[0077] (2) Silicon wafer modification: place the silicon wafer in a volatilization tank, drop 1 to 2 drops of modification reagent HMDS (Hexamethyldisilazane), and volatilize for ≥ 3 minutes;

[0078] (3) Silicon wafer rejection: pour the photoresist on the processed silicon wafer, set the speed of rejection according to the required pattern depth or height, and let it stand for 1 to 2 minutes after uniform glue;

[0079] (4) Silicon wafer exposure: pre-baking process sets the pre-baking time by determining the properties of the photoresist used; the exposure process needs to consider the power of the exposure machine and the metering required for the material to set the exposure time; after exposure, deter...

Embodiment 1

[0095] Prepare a microfluidic chip in the above way (the following microfluidic chips are referred to as chips for short), wherein the size parameters and other parameters are: H1=H2=20 μm, H3=16 μm, H4=16 μm, H5=5 μm, N=0 . The chip was washed with deionized water and phosphate buffered saline (PBS) successively with a syringe pump. Pass the cell carrier fluid into the chip, control the injection speed to 0.1 μl / s, place the chip under a microscope for observation, and finally realize the capture of single cells in the trap. Through Embodiment 1 of the present invention, the capture efficiency of single cells in the trap array reaches 96%, and the single sample consumption is effectively reduced by more than 90% compared with the same concentration of cell culture dishes.

Embodiment 2

[0097] The microfluidic chip was prepared in the above manner, wherein the size parameters and other parameters were: H1=H2=20 μm, H3=16 μm, H4=16 μm, H5=5 μm, N=1. The chip was washed with deionized water and phosphate buffered saline (PBS) successively with a syringe pump. Pass the cell carrier fluid into the chip, control the injection speed to 0.2 μl / s, place the chip under a microscope for observation, and finally realize the capture of single cells in the trap. Through Embodiment 2 of the present invention, the capture efficiency of single cells in the trap array reaches 96%, and the single sample consumption is effectively reduced by more than 95% compared with the same concentration of cell culture dishes.

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Abstract

The invention relates to a micro-fluidic chip for capturing single cells. The chip comprises a function layer and a cover piece layer; the function layer comprises a silicon wafer body and a pluralityof function areas modified on the silicon wafer body; the function areas at least comprise a sample introduction and pretreatment area, a capture function area and a waste liquid treatment area; thesample introduction and pretreatment area is composed of a cell sap inflowing port, an inlet liquid storage tank and an inflowing passage provided with mintype dispersion columns; the capture functionarea is composed of a minitype reaction tank provided with a buffering column and a capture trap array; the waste liquid treatment area is composed of an outflowing passage provided with a minitype dispersion column, an outlet liquid storage tank and a waste liquid outflowing port; the cell sap inflowing port is communicated with the inlet liquid storage tank; the inlet liquid storage tank is communicated with the minitype reaction tank through the inflowing passage; the minitype reaction tank is communicated with the outlet liquid storage tank through the outflowing passage; the outlet liquid storage tank is communicated with the waste liquid outflowing port. The micro-fluidic chip can achieve uniform sample introduction of cell current-carrying liquid and capture of the single cells inthe chip.

Description

technical field [0001] The invention belongs to the field of microfluidic technology, in particular to a microfluidic chip for capturing single cells. Background technique [0002] Since the development of microfluidic technology, it has been widely used in the fields of physical and chemical analysis and biomedicine. Based on micro-nano technology, microfluidic chips provide a good platform for micro-scale research. Due to the similar scale, high analysis efficiency, and high throughput, cell analysis on microchips has many advantages and can be used for medical analysis and drug screening. , body monitoring and other aspects to provide effective research basis and accurate analysis results. [0003] In cell analysis, the analysis of single cells is of great significance in the control of cell morphology and the precise measurement of cells in vivo or in vitro. Different from the tissue analysis of conventional cell groups or cell spheroids, the key research direction of ...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12M1/00
CPCC12M23/16C12M47/04
Inventor 刘坤郝明姜玥陈树雷徐静怡王展威张浩
Owner NORTHEASTERN UNIV
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