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Microfluidic equipment and unit for analyzing single cells and method thereof

A technology of microfluidics and equipment, applied in the field of biology and medical equipment, can solve the problems of device size bubbles and so on

Pending Publication Date: 2022-02-22
SAVELIFE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conventional methods used in the prior art, such as magnetic rotors or blades driven by motors, etc., have problems with device size and the introduction of air bubbles in the liquid

Method used

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  • Microfluidic equipment and unit for analyzing single cells and method thereof
  • Microfluidic equipment and unit for analyzing single cells and method thereof
  • Microfluidic equipment and unit for analyzing single cells and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Embodiment 1 Experimental method and material

[0075] Microfluidic channel preparation:

[0076] Microfluidic channels made of polydimethylsiloxane (PDMS) were fabricated by soft lithography.

[0077] Preparation of UHF Bulk Acoustic Resonator:

[0078] Bulk acoustic wave resonator devices are prepared by chemical vapor deposition, metal sputtering, and photolithography on silicon-based wafers. The specific method is as follows:

[0079] 1. Use a piranha solution with a volume ratio of concentrated sulfuric acid and hydrogen peroxide of 3:1 to thoroughly clean the surface of the silicon wafer. This method can effectively remove organic and inorganic substances on the silicon wafer.

[0080] 2. On the cleaned silicon wafer, a layer of aluminum nitride film is formed by surface sputtering, and then a layer of silicon dioxide film is deposited by ion-enhanced chemical vapor deposition. Then use the same method to alternately deposit aluminum nitride films and silicon ...

Embodiment 2

[0098] In the specific implementation process of this embodiment, a method for analyzing biological particles (such as cells, microvesicles, biological macromolecules such as nucleic acids, etc.) using barcode particles in microfluidic equipment is provided, especially for analyzing the biological particles Nucleic acid methods and microfluidic devices. The method and microfluidic device provided by the present invention can pair single biological particles (such as cells, microvesicles, biomacromolecules such as nucleic acids, etc.) from biological samples with barcode particles, and then combine the paired single biological particles-barcode particles A first liquid (usually an aqueous solution) is contacted with a second liquid (which is a liquid immiscible with the first liquid, usually an oily liquid) to form a droplet containing a paired bioparticle-barcode particle (e.g. water-in-oil droplets). The method also includes further processing and analyzing the nucleic acid ...

Embodiment 3

[0143] Example 3 Sample injection mixing step and sample injection mixing unit

[0144] The method for analyzing biological particles using barcode particles in the microfluidic device provided by the present invention may include the step of sample injection and stirring, and the microfluidic device provided by the present invention may include a sample injection and stirring unit. In the method provided by the invention, a sample containing biological particles (such as cells) to be tested is added into a sample chamber of a microfluidic device, and barcode particles are added into another sample chamber. In one aspect of the present invention, the method comprises stirring the biological sample solutions or particle solutions respectively located in different sample chambers, so as to maintain the dispersion state of biological particles (such as cells) and / or barcode particles (such as magnetic beads), Avoid precipitation and / agglomeration.

[0145] In one aspect of the p...

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Abstract

The invention provides a sample introduction and uniform mixing unit for microfluidic equipment and a sample focusing unit. The invention also provides the microfluidic equipment for analyzing single cell or single nucleic acid by using a bar code particle. The microfluidic equipment is provided with the sample introduction and uniform mixing unit and the sample focusing unit. The invention also provides a method for analyzing the single cell or the single nucleic acid by using the barcode particle.

Description

technical field [0001] The invention relates to the fields of biology and medical equipment. In particular, the present invention relates to a microfluidic system, unit and method for the analysis of single biological particles such as cells or nucleic acids. Background technique [0002] The tool of single-cell sequencing has incomparable advantages in the research of embryos, tumors, and various stem cells. With the continuous breakthrough of single-cell sequencing technology and the widespread use of sequencing instruments, single-cell sequencing analysis has been increasingly applied to various cutting-edge biomedical research. No two cells have genomes that are completely similar at the transcriptome level, so average omics expression analysis of a large number of cells averages out expression signatures that would otherwise be quite prominent in a single cell. [0003] Most of the current single-cell sequencing analysis platforms use microfluidic devices. In microfl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01L3/00C12M1/00C12M1/38C12Q1/6844
CPCB01L3/502784C12Q1/6844B01L2200/12B01L2300/0809B01L2300/0867
Inventor 段学欣杨洋赵海峰
Owner SAVELIFE BIOTECHNOLOGY CO LTD
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