A microfluidic continuous sampling method and device for quantitative analysis of single cells

Abstract

The invention discloses a micro-fluidic continuous sampling method and device for unicellular quantitative analysis. According to the method and the device, a micro-fluidic chip of which cross channels are added with auxiliary focusing channels is used as an operation platform and static pressure provided by samples or electrophoretic buffer solutions in a certain volume ratio in five wide liquid storage tanks at the tail ends of the chip channels and electric power provided by four program controlled direct current voltages are taken as fluid operation control means, thereby realizing parallel treatment of samples and regents in a plurality of microchannels in the micro-fluidic chip, Picoliter-scale injection volume control and continuous injection operation; particularly, the disclosed method and device can be used for realizing three operation steps in one unicellular sample injection to the micro-fluidic chip, wherein the three operation steps include cell sample loading, unicellular loading / capturing and unicellular membrane dissolving and electrophoretic separation; after one unicellular sample injection operation is finished, the three operation steps are repeated automatically and thus the continuous unicellular sample inject is realized.

Description

[0001] Technical field: [0002] The invention belongs to the technical field of single-cell analysis, and relates to a sampling method and device that can provide operations such as "cell loading, single-cell capture / loading, single-cell membrane dissolution, and intracellular component electrophoresis separation" on a microfluidic chip. , especially relates to a microfluidic continuous sampling method and device for quantitative analysis of single cells. [0003] Background technique: [0004] Single-cell quantitative analysis is a new type of experimental technology that realizes single-cell sampling, multi-parameter measurement, and quantitative acquisition of intracellular component information on a single-cell scale. The development and improvement of this technology can not only provide important technical support for the analysis and research of life sciences, medicine and health at the single-cell level and molecular level, but also provide important technical support ...

AI Technical Summary

Problems solved by technology

However, the implementation of this method relies on manual multi-step operations. At the same time, due to its static pressure using a traditional liquid storage tank (that is, the height of the liquid storage tank is greater than its inner diameter), the liquid level of the liquid storage tank will decline with the outflow of the fluid, so that the flow rate It decreases significantly with the drop of liquid level, it is difficult to adapt to the precise control of the injection volume and the continuous injection operation for a long time

Overall, the existing microfluidic single-cell sampling methods still have the following common problems: (1) It is difficult to achieve continuous sampling

After the current sample injection method completes a sample injection and analysis, it usually needs to interrupt the analysis process, and then carry out fluid control means, removal of samples and reagents, cleaning of chips, injection of new samples and reagents, reapplying of control means, etc. A lot of time-consuming operation, causing the analysis throughput of the system to fail to increase

(2) Injection efficiency is low

At present, most of the injection control methods use a simple combination of multiple independent instruments and equipment, and the capture of a single cell often requires the use of a microscope, which makes it difficult to parallelize or synchronize the operations of samples and reagents, which not only leads to poor controllability of the injection volume, but also affects Analyze system stability

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