Chip for open single cell research and preparation method thereof

Abstract

An open single-cell research chip and its preparation method, the chip includes a packaging structure surrounded by a substrate, a retaining wall and a transparent cover, the substrate has a microhole array structure inside the packaging structure, the A number of microwells in the microwell array structure contain single cells respectively, and droplets of reagents for single cell research are respectively added to the top of each microwell by 3D printing, and the remaining space in the packaging structure is filled with sealing oil, further providing The manufacturing method of the chip. The invention perfectly combines 3D printing technology with single-cell microwell arrays, efficiently and quickly realizes the research on the single-cell level, establishes a single-cell research system with simple structure, wide application range and perfect functions, and realizes high occupation High-rate single-cell capture, multi-step liquid treatment of single cells with diversity, accuracy, and open structure can be applied to a variety of research systems.

Description

technical field [0001] The invention relates to the technical field of single-cell research methods, in particular to an open chip for single-cell research and a preparation method thereof. Background technique [0002] Single cell heterogeneity has become one of the key research objects in the field of cell biology. However, since the size of single cells is about 15 μm, the capture, screening, sample processing and signal detection of single cells pose great challenges to traditional experimental methods. Microfluidic technology is expected to become a powerful tool for single-cell research as a means of precisely and efficiently handling picoliter-level fluids. At present, a variety of new microfluidic chips have been successfully applied to single-cell level research, but these methods still have limitations. [0003] The first type is single-cell microwell structure arrays to capture single cells. This type of single-cell microwell can only accommodate one cell, so the...

AI Technical Summary

Problems solved by technology

At present, a variety of new microfluidic chips have been successfully applied to single-cell level research, but these methods still have limitations

[0003] The first type is single-cell microwell structure arrays to capture single cells. This type of single-cell microwell can only accommodate one cell, so the occupancy rate of single cells can break through the limit of Poisson distribution and can reach 97%. However, the space of micropores is small. It is difficult to provide enough reagents for subsequent detection experiments, making it difficult for the single-cell microwell structure array chip to carry out the later cell processing process

[0004] The second type is the traditional T-channel water-in-oil microfluidic device. This type of microfluidic device can quickly and high-throughput generate microdroplets containing a small number of cells. However, there is an insurmountable contradiction in this method: if the droplet If the volume is large, the number of single cells obeys the Poisson distribution; if the volume of the droplet is small, the amount of reagent in a single droplet is not enough

[0005] The third type of microfluidic device can realize the capture of single cells and subsequent liquid injection by designing complex microstructures and coordinating with the switch control of the valve. An open approach to single-cell research

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