High-flux cell electrofusion device based on microporous array film

Abstract

The invention discloses a high-flux cell electrofusion device based on microporous array film, which comprises a microporous array film, two sample cells and a cover plate, wherein the two sample cells are respectively attached to both surfaces of the microporous array film, and the cover plate is arranged on the microporous array film and two sample cells; the middle part of the microporous array film is provided with a microporous array; and the lower ends of the two plate electrodes respectively penetrate through the cover plate and are inserted into the two sample cells. A microporous array film with massive micropores is processed and integrated by micro-processing technology, the microporous structure is utilized to change the electric field distribution between the two plate electrodes, and thus, the micropore central region is the highest electric field region; the dielectrophoresis effect can be utilized to control cells in the sample cells to move towards the high electric field region; both sides of each micropore respectively adsorb one cell, and the center of each micropore forms cell pairing and connection; the micropore center has the highest electric field intensity, and the maximum fusion probability, thereby avoiding multicellular fusion; and the high-flux cell electrofusion device has favorable biocompatibility and corrosion resistance, thereby ensuring the reliability of the device and enhancing the safety of cell fusion.

Description

technical field [0001] The invention relates to a high-throughput cell electrofusion device based on a microporous array film, which is mainly used in the preparation of fusion cells of animals, plants, and microbial cells, and can also be used in researches such as electroporation and electrotransfection. It can be widely used in genetics, animal and plant distant hybrid breeding, developmental biology, drug screening, monoclonal antibody preparation, mammalian cloning and other fields. Background technique [0002] Since the 1980s, cell electrofusion technology has been rapidly developed and widely used because of its high efficiency, simple operation, non-toxicity to cells, easy observation, suitable for instrument application and standardized operation. Cell electrofusion can be divided into two main stages: cell queuing and cell fusion. [0003] The principle of cell queuing is that when biological cells are in a non-uniform electric field, they are polarized by the el...

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Problems solved by technology

[0010] (1) Due to the large distance between the electrodes in the fusion tank, to achieve sufficient strength of cell queuing, fusion and compaction signals, a high external driving voltage is required, often as high as hundreds of thousands of volts, which requires the electrical safety of the system High, the cost of the system is also greatly increased;

[0011] (2) The large distance between electrodes is not conducive to the precise control of cells, etc.

[0013] However, there are still certain problems with the above-mentioned chips. For example, although the microfluidic chip researched by MIT in the United States has solved the problem of cell pairing, the distance between the two electrodes of the chip is relatively large, and a high external voltage is still required to be able to achieve electrical fusion

However, the number of microelectrodes integrated in the chip proposed by Zhao Zhiqiang is small, and high-throughput fusion cannot be achieved; the electric field strength and electric field gradient generated by the microelectrodes are relatively weak, and it is difficult to achieve precise control of cells; the corrosion resistance of the selected processing materials , the anti-oxidation ability is also poor; at the same time, because the sample inlet and outlet device is not integrated, the operation is also more inconvenient

Japanese researchers proposed to use micropores to separate the two channels, and then achieve independent injection of different cells to complete cell pairing. However, this method has high requirements for the positioning of micropores and cannot be applied on a large scale. Still use the plate electrodes with a large distance to achieve electrofusion, and the working voltage is high

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