Single cell capturing system based on image analysis

Abstract

The invention discloses a single cell capture system based on image analysis. The single cell capture system comprises a liquid storage unit, an optical imaging unit, an image analysis unit, a sampling unit and a control unit which is respectively connected with the sampling unit and the image analysis unit, the optical imaging unit is used for acquiring image information in the liquid storage unit and comprises a light source and an image acquisition element; the sampling unit comprises a sampling suction head and a sampling pipeline which are communicated; the sampling suction head sucks a target object from the liquid storage unit and transfers the target object through the sampling pipeline communicated with the sampling suction head; the image analysis unit is connected with the image acquisition element to receive and output image information; the control unit comprises a first displacement control module for controlling the sampling suction head; the first displacement control module is connected with the image analysis unit; and the first displacement control module controls the displacement of the sampling suction head according to information fed back by the image analysis unit. The single cell capture system provided by the invention can realize capture of single cells and single cell analogues with high efficiency and high specificity.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to a single cell capture system based on image analysis. Background technique [0002] Single-cell sequencing technology refers to the quantitative sequencing of the genome, transcriptome and other information of a single cell, thereby revealing cell population differences and cell evolutionary relationships. High-throughput sequencing technology is Next Generation Sequencing (NGS), which can conduct a comprehensive and detailed analysis of the genome and transcriptome of a species, and can perform parallel analysis of hundreds of thousands to millions of DNA molecules at a time. Sequence determination, short read length, etc. are marked by reading multiple short DNA fragments and splicing into complete sequence information; it has significant advantages in processing large-scale samples, and is the core technology in current omics research. [0003] In 1964, Cornell...

AI Technical Summary

Problems solved by technology

[0012] 1) The single-cell technology based on microfluidics has a clear limit on the size of the cells. The microfluidic chip of 10x Genomics generally requires the diameter of the cells to be less than 40um. Cells larger than 40um will cause problems such as sample preference and loss of sample information

At the same time, it is difficult to capture too small cells or cell-like cells, such as organelles, etc., which limits the application range of the technology;

[0013] 2) Since the current mainstream method uses the form of water-in-oil droplets to form a micro reaction system for nucleic acid amplification, this method has a certain preference for cell types, such as adipocytes are less efficient in the process of forming water-in-oil droplets Then affect the efficiency of library construction and amplification, etc.

Since the nuclear and cytoplasmic separation of single cells cannot be achieved, single-cell multi-omics library construction and sequencing cannot be achieved;

[0015] 4) The current mainstream microfluidic and Microwell technology systems, due to the limitations of capture technology and limited sequencing depth, etc., the number of genes that can be detected in the average single cell finally obtained is small, so part of the sample information may be lost and it is not conducive to the depth of the later stage. data mining analysis

[0016] 5) The current mainstream microfluidic and Microwell technology systems capture cells indiscriminately, in order to improve the capture efficiency of living cells and produce higher-quality sequencing data, so the current capture technology systems require the loading of single-cell suspensions. The cell viability is tested first, and the cell viability in the suspension must reach a certain standard. If the cell viability is above 85%, then there is no difference in capture. For the captured apoptotic cells, the follow-up can only be done through bioinformatics algorithm for filtering

[0017] 6) The current mainstream microfluidic and Microwell technology systems adopt the strategy of mixing cells after capture to build an overall library. Due to the limitation of capture efficiency, it requires a large number of cells to be loaded, at least hundreds of cells, and generally requires Tens of thousands of cells, so it is not suitable for samples with rare cells, such as single-cell samples of single embryos, etc.

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