High-flux cell electrical fusion microfluidic chip device based on microporous microelectrode array

Abstract

The invention discloses a high-flux cell electrical fusion microfluidic chip device based on a microporous microelectrode array, which consists of a microelectrode microporous array chip and a microfluidic cover plate with top electrodes. In the invention, a micro electro mechanical system (MEMS) processing technique is adopted, and a microporous microelectrode array is constructed on a quartz substrate by using both a microelectrode array and a polymer microporous array; the microporous microelectrode array can better control cell pairing, so that a pair of cells fuses in each micro pore; the array structure can improve the flux of cell fusion; and the materials used for making the device disclosed by the invention have high biocompatibility and corrosion resistance and can ensure the reliability of chip and improve the safety of cell fusion. The device can be widely used in fields of genetics, animal and plant distant hybridization breeding, development biology, medicine screening, preparation of monoclonal antibody, mammalian cloning and the like.

Description

technical field [0001] The invention relates to a device for electrical fusion of biological cells. Specifically, the present invention relates to a chip for providing cell electrofusion, providing and generating the electric field intensity and electric field gradient required for cell alignment, electroporation, and fusion. Background technique [0002] Since the 1980s, cell electrofusion technology has been rapidly developed and widely used because of its advantages such as high efficiency, easy operation, non-toxicity to cells, easy observation, suitable for instrument application and standardized operation (see US patent literature : 4326934, April 27, 1982, Pohl; 441972, April 10, 1982, Pohl; 4578168, March 25, 1986, Hofman; 4695547, Sep 22, 1987, Hillard; 4699881, Oct 13, 1987, Matschke et al. ;). [0003] Cell electrofusion can be divided into two main stages: cell queuing and cell fusion. [0004] The principle of cell queuing is that when biological cells are in...

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Problems solved by technology

[0007] However, the traditional cell electrofusion equipment also has some disadvantages: (1) Due to the large electrode spacing in the fusion tank, a high external driving voltage is required to achieve sufficient strength of cell queuing, fusion and compaction signals, often as high as several Hundreds of thousands of volts, high requirements on the electrical safety of the system, the cost of the system is also greatly increased; (2) The large distance between electrodes is not conducive to the precise control of cells, etc.

However, the electrode in this patent is a continuous electrode, and the side wall of the microchannel in the depression area is also an electrode structure. The electrical signal loaded on it will also cause the cells to line up, but the electric field strength in this area is not enough to achieve fusion, which will affect the efficiency (Chinese Patent Document 200710092892.2 )

At the same time, the cell pairing of the cell electrofusion chip device based on the micro-chamber array structure proposed by the research group is based on a random principle of cell pairing and fusion. Although it can achieve the goal of pairing two cells, it still cannot accurately realize the fusion of AB cells. Pairing and Fusion (Chinese Patent Document 200910191052.0)

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