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Method for processing cell sample in single cell sorting process

A sorting process and processing method technology, applied in cell dissociation methods, microorganism-based methods, plant cells, etc., can solve the problems of difficult separation and acquisition of cultured microorganisms, restrict the growth of micro flora, etc., and achieve the goal of maintaining cell activity. Effect

Inactive Publication Date: 2020-12-15
长春长光辰英生物科学仪器有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many kinds of environmental microorganisms, and the total number is huge. Different microorganisms have different requirements for energy, nutrition and physical and chemical conditions. The existing dilution coating plate method and plate streaking method are difficult to separate microorganisms that are difficult to cultivate. Acquired, and limited growth of many minor flora in complex mixed samples due to growth inhibition of dominant flora

Method used

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  • Method for processing cell sample in single cell sorting process
  • Method for processing cell sample in single cell sorting process
  • Method for processing cell sample in single cell sorting process

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Embodiment 1

[0029] Preferred embodiment 1 of the present invention provides a method for processing cell samples in the single cell sorting process, wherein the cell samples are microbial cell samples, specifically including the following steps in order:

[0030] Step 1. Obtain the required cell samples. The specific operations are as follows:

[0031] Soil sample culture: the soil sample is obtained from the environment, and then placed in the medium for culture, and the culture time is about 10 days;

[0032] Sample preparation: ①Collect 1ml of the above cultured soil sample and filter through a 40μm filter membrane to remove impurities in the soil. ②Centrifuge at 5000rpm for 5min to obtain fine bacterial precipitate. ③Add 1ml of sterilized ultrapure water to the bacterial pellet, wash 5 times, centrifuge at 5000rpm for 5min after each wash, and collect the bacterial pellet. ④Add 1ml of ultrapure water to shake and mix well to obtain microbial cell samples for later use.

[0033] Ste...

Embodiment 2

[0044] Preferred embodiment 2 of the present invention provides a method for processing cell samples in the single cell sorting process, the cell samples are Bacillus subtilis cell samples, specifically including the following steps in order:

[0045] Step 1. Obtain the required cell samples. The specific operations are as follows:

[0046] Sample culture: inoculate the sample to be separated in LB medium, and culture overnight at 37°C with shaking.

[0047] Sample processing: ①Collect 1ml of overnight cultured bacterial solution in a 1.5ml centrifuge tube, centrifuge at 5000rpm for 5min to obtain bacterial precipitates. ②Add 1ml of sterilized ultrapure water to wash 4 times, centrifuge at 5000rpm for 5min after each wash, and collect the bacterial precipitate. ③ Add 1ml of ultrapure water, shake and mix well, and obtain cell samples for later use.

[0048] Step 2: Spread the cell sample and the water-retaining substance used to maintain cell activity on the surface of the s...

Embodiment 3

[0058] Preferred embodiment 3 of the present invention provides a method for processing cell samples in the single cell sorting process, the cell samples are microbial cell samples, specifically including the following steps in order:

[0059] Step 1. Obtain the required cell samples. The specific operations are as follows:

[0060] Soil sample culture: the soil sample is obtained from the environment, and then placed in the culture medium for culture, the culture time is about 15 days;

[0061] Sample preparation: ①Collect 1ml of the above cultured soil sample and filter through a 40μm filter membrane to remove impurities in the soil. ②Centrifuge at 5000rpm for 5min to obtain fine bacterial precipitate. ③Add 1ml of sterilized ultrapure water to the bacterial pellet, wash 3 times, centrifuge at 5000rpm for 5min after each washing, and collect the bacterial pellet. ④Add 1ml of ultrapure water to shake and mix well to obtain microbial cell samples for later use.

[0062] Step...

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Abstract

The invention provides a method for processing a cell sample in a single cell sorting process. The method comprises the following steps in sequence: 1, acquiring a required cell sample; 2, spreading the cell sample and a water retention substance for keeping cell activity on the surface of a sorting chip; 3, putting the chip on a sorting platform for sorting target cells; 4, putting the sorted cells into a receiving substance in a receiving device; and 5, performing subsequent detection or culture operation on the sorted cells. According to the method, the substance with a water retention function is used, so that the cells can keep the original biological state and biological activity in the sorting process, and detection and enlarged culture of the activity of the cells after sorting arenot influenced.

Description

technical field [0001] The invention relates to a cell sample processing method, in particular to a cell sample processing method in the single cell sorting process. Background technique [0002] Cells are the basic unit of life. Due to the phenotypic heterogeneity of cells, studying them at the single-cell level is of great significance for analyzing the deep operating mechanism of life activities. [0003] At present, only a very small number of microorganisms in nature can be isolated and cultivated, which seriously hinders the research on the laws of microbial life activities and the development of microbial resources. [0004] In order to promote the study of microbial diversity, we need to isolate and culture microbes that are difficult to culture from complex samples. However, there are many kinds of environmental microorganisms, and the total number is huge. Different microorganisms have different requirements for energy, nutrition and physical and chemical conditio...

Claims

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Application Information

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IPC IPC(8): C12N1/02C12N5/04C12N5/07C12R1/125
CPCC12N1/02C12N5/00C12N5/04C12N2509/10
Inventor 薛莹李航顾珈瑜田春悦李备
Owner 长春长光辰英生物科学仪器有限公司
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