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System and method for low-cost and efficient separation and obtaining of single cell

A technology for separation and acquisition of single cells, applied in the fields of biotechnology and instrument science, which can solve the problems of uncontrollable separation and acquisition of single cells, high price, and low separation throughput, and achieve low cost, short time-consuming, and scalability strong effect

Active Publication Date: 2015-09-02
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The first two technical platforms are expensive and have low separation throughput; the infinite dilution method is simple and convenient, but cannot be controlled to separate and obtain single cells

Method used

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  • System and method for low-cost and efficient separation and obtaining of single cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Low-cost and high-efficiency separation and acquisition of single cells based on routine laboratory imaging techniques

[0040] Such as figure 1 , 2 As shown, the low-cost and high-efficiency system for separating and obtaining single cells based on conventional laboratory imaging technology constructed in this example, the structure of the microfluidic chip 7 is shown in figure 1 , which includes: 1. Continuous oil phase inlet; 2. Cell suspension inlet; 3. Side flow oil phase inlet; 4. Electromagnetic valve connection port; 5. Waste liquid outlet; 6. Sorted droplet outlet. The microfluidic chip 7 is formed by bonding the upper layer of PDMS and the lower layer of glass substrate, wherein the upper layer has a 25-150 micron wide and 40-60 micron deep microchannel network obtained by wet etching, and the lower glass substrate is attached with a A layer of PDMS with a thickness of about 1 mm is used to easily produce water-in-oil droplets and bond seals betwe...

Embodiment 2

[0041] Example 2: Efficient separation and acquisition of single cells based on high-speed camera and image processing technology

[0042] Such as figure 1 , 2 As shown, the system constructed in this example is based on a high-speed camera and image processing method to efficiently separate and obtain single cells. The structure of the microfluidic chip 7 is shown in figure 1, which includes: 1. Continuous oil phase inlet; 2. Cell suspension inlet; 3. Side flow oil phase inlet; 4. Electromagnetic valve connection port; 5. Waste liquid outlet; 6. Sorted droplet outlet. The microfluidic chip 7 is formed by bonding the upper layer of PDMS and the lower layer of glass substrate, wherein the upper layer has a 25-150 micron wide and 40-60 micron deep microchannel network obtained by wet etching, and the lower glass substrate is attached with a A layer of PDMS with a thickness of about 1 mm is used to easily produce water-in-oil droplets and bond seals between the upper and lower ...

Embodiment 3

[0043] Example 3: Low-cost and efficient separation and acquisition of single cells based on fluorescence detection technology

[0044] Such as figure 1 , 2 As shown, the low-cost and high-efficiency separation and acquisition system based on fluorescence detection technology constructed in this example, the structure of the microfluidic chip 7 is shown in figure 1 , which includes: 1. Continuous oil phase inlet; 2. Cell suspension inlet; 3. Side flow oil phase inlet; 4. Electromagnetic valve connection port; 5. Waste liquid outlet; 6. Sorted droplet outlet. The microfluidic chip 7 is formed by bonding the upper layer of PDMS and the lower layer of glass substrate, wherein the upper layer has a 25-150 micron wide and 40-60 micron deep microchannel network obtained by wet etching, and the lower glass substrate is attached with a A layer of PDMS with a thickness of about 1 mm is used to easily produce water-in-oil droplets and bond seals between the upper and lower layers. Dr...

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Abstract

According to the present invention, based on a liquid droplet microfluidic chip technology, a system and a method for low-cost and efficient separation and obtaining of single cell is designed and constructed. The system comprises a microfluidic chip, a liquid droplet signal detector, a signal analysis and sorting control unit, an electromagnetic valve, a conduct pipe, and a liquid droplet collection micro-pipe. The single cell separation process comprises that: liquid droplets wrapping cells are subjected to online continuous generation and directional flow, signal acquisition and analysis and sorting control are performed when passing through a signal acquisition zone, and when the sorting condition (for example: the liquid droplet only warps single cell) is met, the work of the electromagnetic valve is controlled to make the force applying state and the flow direction of the liquid droplet be changed so as to separate and collect the single liquid droplet, such that the obtaining of the single cell can be achieved through the de-emulsification, the extraction and other operations. According to the technical method of the present invention, a set of the simple and feasible system and the simple and feasible method suitable for the liquid droplet sorting, cell separation and single cell obtaining of different detection methods are provided, and the method has advantages of high efficiency, short time consumption, low cost, no pollution, wide application range, strong extensibility and the like compared with the traditional single cell obtaining method.

Description

technical field [0001] The invention belongs to the fields of biotechnology and instrument science, and specifically relates to a system and method for realizing low-cost and high-efficiency separation and acquisition of single cells based on the methods of droplet wrapping and droplet sorting on a microfluidic chip. Background technique [0002] Living single cells are the basic unit of life activities and evolution. It is of great significance to study cell biological processes at the level of single cells. It can reveal the pathogenesis of cancer in essence, understand the principles of cell differentiation and tissue development, and identify gene expression characteristics. and cell characteristics. [0003] Rapid isolation and acquisition of single cells has become a key technology for single cell research and analysis. Flow cytometry is an important instrument platform for single cell sorting and analysis, and is an effective means of separating cells. However, sinc...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12N1/18C12R1/865
CPCC12M23/16C12M41/48C12M47/04
Inventor 马波张强徐健
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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