Integrated micro-fluidic chip and system for capture, culture and administration of single cells

Abstract

Aiming at the lack of an integrated micro-fluidic chip capable of realizing capture, culture and batch differential administration of single cells simultaneously in the prior, the invention provides an integrated micro-fluidic chip and system for capture, culture and administration of single cells and belongs to application of micro-fluidic chip technologies to the field of cell pharmacology analysis. The integrated micro-fluidic chip comprises a cell capture and culture unit, a drug diffusion unit and a drug supply unit, which are sealed in sequence from top to bottom; the system employing the integrated micro-fluidic chip comprises the integrated micro-fluidic chip, a fluorescence microscope, image processing equipment and a digital injection pump. A preparation method of the integrated micro-fluidic chip comprises the following steps: photo-etching baseplate plane figures on the silicon plates of the three units to etch grooves, pouring a liquid chip material into the grooves, and obtaining the three units after the liquid chip material is solidified; conducting sealing bonding and punching to obtain the integrated micro-fluidic chip. The integrated micro-fluidic chip provided by the invention integrates the three functions of capture, culture and batch differential administration of single cells and effectively avoids sample contamination.

Description

technical field [0001] The invention belongs to the application of microfluidic chip technology in the field of cell pharmacological analysis, and in particular relates to an integrated microfluidic chip and system for single cell capture, cultivation and drug delivery. Background technique [0002] In today's society, various diseases, especially cancer, seriously threaten human health. Single-cell level research is very important for the research of major diseases such as cancer, blood diseases, Parkinson's disease, and Alzheimer's disease. The growth, metastasis, and spread of cancer cells and other cells, as well as postoperative judgment, are of great significance. [0003] In recent years, with the rapid development of micro-electromechanical technology, life science, analytical chemistry and other disciplines, bio-MEMS technology and micro-total analysis system (u-TAS) based on Bio-MEMS technology are gradually becoming the research hotspots of scientists. The micro...

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Problems solved by technology

[0004] In Lab Chip, 2006, 6, 1445–1449, Dino Di Carlo et al. used soft lithography technology and PDMS materials to design and manufacture a multi-array single-cell capture device with a U-shaped microwell structure in a single direction. The device achieves rapid capture of single cells; however, the researcher did not specify the long-term culture after cell capture, batch differential drug delivery, etc.

[0005] In Nature Methods, 2009, 6(2), 147-152, Alison M Skelley et al. used soft lithography technology and PDMS materials to design and manufacture a multi-array cell with a U-shaped microwell in each direction. Capture, pairing, and fusion device. This device is combined with fluorescent tracking technology to observe the dynamic whole process of pairing and fusion of captured cells in the U microwell; Studies on drug administration, etc. are not specified

[0006] In Sensors and Actuators A.215(2014), 197-203, Jing Zhu et al. adjusted the geometric structure and stress distribution of the microwell in the PDMS microfluidic chip, and the control of the microwell in the single-cell or multi-cell capture chip. Regulation was studied; however, this investigator did not study bulk differential dosing after single cell capture

[0007] Patent application number CN201410141283.1: A microfluidic chip and system for single-cell analysis and a single-cell analysis method. A chip system is designed to realize single-cell capture, lysis, and intracellular component analysis; but the integrated chip The capture of single cells is achieved by setting vertical capture holes, driven by gravity, the process is not easy to manipulate and is easy to cause damage to cells; functionally, the chip cannot realize the cultivation and batch differential drug delivery after single cell capture

[0008] Patent application number CN201110224663.8: a whole blood cancer cell capture integrated microfluidic chip, designed a microfluidic chip that can separate and capture whole blood cancer cells; the microfluidic chip is a one-layer unit structure, It can achieve the separation and capture of multiple cells, but cannot achieve the capture, culture and batch differential drug delivery of single cells

[0009] The patent application number is CN201180072343: microfluidic cell capture and analysis equipment for high-throughput analysis, a microfluidic chip unit for multi-cell capture and analysis is designed, but the disadvantage is that the chip is in the design size and In principle, the structure cannot ensure the capture of single cells

[0010] The patent with the publication number US8475730B2: Apparatus for single cell separation and position fixing, designed an integrated microfluidic chip with a three-layer unit structure, but the function of the chip is mainly to separate and position a single cell. The defect is that diffusion micropores and drug delivery chambers can no longer be set on this structure, so batch differential drug delivery to single cells cannot be realized

[0011] The patent with the publication number US005821073A: Method and apparatus for single step assays of whole blood, designed a three-layer unit structure integrated microfluidic chip, but the main function of the integrated chip is to analyze the cell mass and other substances in the blood. Separation analysis, the separation is not single cells, but large-scale clusters such as multi-cell clusters and some protein molecular clusters; the design scale and capture structure cannot capture single cells

[0012] Patent publication number WO2014066888 (A1): High-Throughput Mutagenized Cell Screening System For Selective Single Cell Extraction, an integrated microfluidic chip with multi-layer units is designed, but the main function of the integrated chip is to screen single cells , to extract cell substances, and the design of the internal channel of the integrated chip makes it unsuitable for single cell culture and batch differential drug delivery

[0013] Among the technologies that have been disclosed so far, there are very few literatures related to the subsequent cultivation and administration of captured single cells. At present, there is no design and manufacture of an integrated microfluidic chip to realize the capture, cultivation and batch production of single cells at the same time. Reports of differential dosing

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