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Pressure sucking-based cell/particle sorting system and method

A sorting system, cell technology, applied in the field of biotechnology and instrument science, can solve problems such as restriction and disturbance of fluid flow pattern

Inactive Publication Date: 2015-06-03
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

In addition, there is a switch to control the external pressure through the solenoid valve to realize the deflected flow pattern of the cells / particles to be sorted into the sorting channel. This method is relatively simple and easy to implement. There is an insurmountable defect in the particle deflection method, that is, when the solenoid valve is opened, the suction of the liquid will disturb the flow pattern of the fluid in the microchannel, especially when precise control of the entrainment is required to achieve single-cell flow. Applications in point-of-check cell / particle sorting systems are limited

Method used

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  • Pressure sucking-based cell/particle sorting system and method

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Example 1: Raman activated single cell sorting based on pressure sucking

[0033] Such as figure 1 , 2 As shown, the quartz microfluidic chip constructed in this example is used for single-cell sorting based on pressure-sucking Raman activation in the flow state. The chip structure is shown in figure 1 , including 1. Precision micropump or static pressure driven entrainment buffer inlet; 2. Precision micropump or static pressure driven cell suspension inlet; 3. Waste liquid outlet; 4. Sorted cell / particle outlet for The liquid conduit 10 is connected with the solenoid valve 9; 5, 6, the electrode pair are connected with the output of the high-frequency AC signal generator. The chip is composed of upper and lower quartz substrates. The cover sheet includes a microchannel network of 20-150 microns wide and 10-50 microns deep by wet etching. The micro-electrode structure on the lower quartz substrate is obtained by photolithography and electrode etching solution. etched...

Embodiment 2

[0034] Example 2: Fluorescence cell sorting based on pressure suction

[0035] The microfluidic chip constructed in this example is used for fluorescent cell sorting based on pressure suction in a flowing state. Outlet 4 is the same as the pressure deflection control port. The microfluidic chip 12 is composed of upper and lower substrates, the upper substrate includes a microchannel network with a width of 20-150 microns and a depth of 10-50 microns, and the lower layer is a cover sheet. The upper and lower substrates are cleaned, aligned precisely under a microscope, and then bonded. The outlet and inlet of the chip are connected to the outside world through fixed interfaces and microtubules to realize the introduction and separation of cells / particles. The microfluidic chip 12 and related methods described in this example realize the sorting of single cells based on pressure sucking Raman activation in a flow state. Cells (expressed with fluorescent protein) are diluted t...

Embodiment 3

[0036] Example 3: Sorting of microdroplets based on pressure suction

[0037]The microfluidic chip 12 and method constructed in this example are used for sorting fluorescent micro-droplets based on pressure sucking in a flowing state. The chip structure includes: 1. Oil phase inlet (entrainment buffer inlet); 2. Droplet inlet (cell suspension inlet); 3. Waste liquid outlet; 4. Sorting droplet outlet (sorting cell / particle outlet), the droplet collection port is the same as the pressure deflection control port. The chip is composed of upper and lower substrates, the upper substrate includes a microchannel network with a width of 20-150 microns and a depth of 10-50 microns, and the lower layer is a cover sheet. The upper and lower substrates are cleaned, aligned precisely under a microscope, and then bonded. The outlet and inlet of the chip are connected to the outside world through fixed interfaces and microtubules to realize the introduction and separation of cells / particles....

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Abstract

According to a solenoid valve switch principle comprising that at the solenoid valve opening moment, liquid in a liquid channel is instantly sucked, a pressure sucking-based flowing cell / particle sorting system and method are designed and constructed by effective utilization of sucking effects. The system and method effectively solve the problems of the pressure driving-based cell / particle sorting system and improve sorting accuracy and efficiency. The sorting system comprises a cell signal acquisition and analysis unit, a micro-fluidic chip and a pressure control unit. The continuously-flowing cell / particle flows flow in a micro-fluidic chip micro-channel under the power action, when the cells flow through a detection point of the cell signal acquisition and analysis unit, a cell signal is acquired and determined, and when the specific cells satisfying sorting conditions are detected, the determined signal activates the pressure control unit, and the pressure control unit sucks the specific cells / particles so that the specific cells / particles enter into the cell / particle sorting channel.

Description

technical field [0001] The invention belongs to the fields of biotechnology and instrument science, and specifically uses the principle of pressure suction to construct a system and method for separating specific cells / particles from groups in a flowing state. Background technique [0002] Cells are the basic unit of organism structure and function. The study of individual differences in single cells can reveal the pathogenesis of cancer, understand the principles of cell differentiation and tissue development, and identify gene expression characteristics and cell characteristics. [0003] Rapid identification and isolation of single cells with specific functional phenotypes has become a key technology for single cell analysis. Flow cytometry is an important instrument platform for single cell sorting and analysis, and is an effective means of separating cells. However, since most living cells have weak or no fluorescence, the addition of fluorescent markers is not suitable...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/10C12M1/42C12M1/36C12M1/34C12N5/00
Inventor 马波张沛然徐健张旭
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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