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Method for isolating single cells

A single cell and cell technology, applied in the field of single cell separation, can solve the problems of easy inactivation of cells and low separation efficiency, and achieve the effect of improving separation accuracy, improving separation efficiency and maintaining activity.

Active Publication Date: 2017-05-31
GUANGDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In view of this, the object of the present invention is to provide a method for separating single cells, which is used to solve the problems of low separation efficiency of existing single cell separation techniques and easy inactivation of cells during the separation process

Method used

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  • Method for isolating single cells
  • Method for isolating single cells
  • Method for isolating single cells

Examples

Experimental program
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Embodiment 1

[0046] In this embodiment, yeast cells are taken as an example, and the specific steps for single cell isolation are as follows:

[0047] 1. Put the experimental equipment into a high-pressure steam sterilizer, and sterilize in high-pressure steam at 121°C for 20 minutes.

[0048] 2. When yeast cells grow in cell culture medium, the position of the cells is not fixed, which easily affects the separation accuracy of single cells. For this reason, in this experiment, 5% polylysine (PLL) was used to treat one side of the photolysis material layer, so that the yeast cells could not swim freely.

[0049] 3. Coat the surface of the photolytic material layer with a thickness of about 40 μm and a cell density of 3×10 by scraping coating technology. 6 cells / mL of cell culture medium, and then placed at 37°C, 5% CO 2 Stand still in the cell incubator for 30 minutes, so that the yeast cells land and fix on the surface of the photolysis material layer.

[0050] 4. Use a 532nm nanosecon...

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Abstract

The invention relates to the technical field of single-cell isolation, in particular to a method for isolating single cells. With the method, size of single droplets can be accurately controlled by regulating size of laser energy, diameter of laser spots and thickness of photolytic material layers, only one cell is wrapped in each droplet, and isolation accuracy of the single cells is greatly improved; in addition, the laser energy does not act on the target cells direction, so that incompletion of cell functions cannot occur, and activity of the cells is sustained; meanwhile, the cells do not needed to be marked with fluorescent protein or magnetic particles, manual operation is omitted, and isolation efficiency is greatly improved.

Description

technical field [0001] The invention belongs to the technical field of single cell separation, and in particular relates to a method for separating single cells. Background technique [0002] Single cell technology takes single cells as the research direction to study the growth, metabolism and apoptosis of single cells. Recently, single cells have been widely used in biomedical fields such as personalized therapy, tumors, cardiovascular and cerebrovascular diseases, fertility and AIDS. Single cell isolation technology can provide research samples for single cell research, which is a crucial step in the realization of single cell technology. [0003] Traditional methods for isolating single cells mainly include limiting dilution, fluorescence-activated cell sorting (FACS), magnetic-activated cell sorting (MACS) and optical tweezers. The effective dilution method is mainly manual operation, the process is complicated, and the separation efficiency is low; the fluorescence a...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12N1/02
CPCC12N1/02C12N1/16
Inventor 邓宇王文兵郭钟宁张永康黄志刚刘江文朱紫红麦文豪洪文生
Owner GUANGDONG UNIV OF TECH
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