Micro-fluidic chip for capturing single cell/single granule through high-flux pairing and application of micro-fluidic chip

Abstract

The invention relates to a micro-fluidic chip for capturing single cells / single granules through high-flux pairing and application of the micro-fluidic chip. The micro-fluidic chip comprises three parts, namely a capturing layer, a control layer and a carrier from top to bottom, wherein the capturing layer consists of two runners in parallel and a connection channel; the control layer is an isolation pump layer; a plurality of single granule / single cell capturing units are arranged inside the runners and are used for capturing single granules / single cells; the connection channel is used for communicating two paired capturing units; isolation channels are positioned below the connection channel, are perpendicular to the connection channel and are isolated by diaphragms. The micro-fluidic chip is capable of capturing single granules / single cells in a high flux manner, is high in capturing efficiency, capable of achieving pairing of single granules / single granules, single cells / single cells and single granules / single cells, high in pairing efficiency, capable of achieving parallel operation and control on paired units, such as isolation, culture, cracking and nucleic acid amplification of single cells, can be widely applied to the field of single cell analysis, and is applicable to cell samples of small numbers.

Description

Technical field [0001] The invention relates to a microfluidic chip, in particular to a microfluidic chip with high-throughput paired capture single cells / single particles. Background technique [0002] Cells are the most basic unit of life body composition and life activities. The traditional method of analyzing and processing the average signal of a large number of cells makes the averaging of the signal obscure people's understanding of the heterogeneity of the brain, blood system, immune system, and the cells that make up these systems. With the development of high-throughput sequencing technology, single-cell sequencing technology has become one of the most important methods for single-cell analysis, which greatly improves the efficiency and accuracy of single-cell analysis. Analysis of single-cell gene levels can reveal mutations and structural variations in the genome of cells (especially cancer cells), understand the differences in response to drugs between cells, and fi...

AI Technical Summary

Problems solved by technology

[0005] However, existing microfluidic chips cannot achieve single-cell capture of a small number of rare cells, and cannot perform high-throughput analysis on a large number of single cells at the same time.

For example, in the drop-seq method reported in the Cell article (Macosko et al., 2015, Cell: 161, 1202-1214), the capture of single cells is based on the Poisson distribution principle, and most of the droplets have no cells, only ~1% The droplet contains a single cell, combined with the Poisson distribution of the microsphere, the effective analysis target is further reduced, and only a small number of cells in a large number of actual samples can be analyzed, which may ignore some important individual cells in the sample

In addition, the drop-seq method is only suitable for analyzing samples with a large number of objects. For some rare cells (such as circulating tumor cells), due to the small number of cells in the sample (10-100 / mL blood), this method cannot be used. single cell analysis

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