Cell electrofusion chip device based on columnar microelectrode array and electrofusion method

Abstract

The invention provides a cell electrofusion chip device based on a columnar microelectrode array, which consists of a columnar microelectrode array chip, a printed circuit board and a flow passage control module. The columnar microelectrode array chip consists of a quartz substrate layer, a metal lead layer, a polymer insulated layer and a columnar microelectrode layer sequentially from bottom totop, wherein the metal lead layers of the columnar microelectrode array chip is electrically connected with the periphery printed circuit board in a bonding way, leads outside electrical signals ontoa columnar microelectrode, and leads a gradient electric field with enough strength to be formed between adjacent columnar microelectrodes; and the flow passage control module covers the columnar microelectrode array chip. In the invention, by loading electrical signals on the columnar microelectrode, queue and fusion among cells are controlled; simultaneously, the superiority that the columnar microelectrode array benefits the flow of the cells in chamber-tracts is utilized for realizing continuous stream fusion of the cells; and with the combination of the design of the array columnar microelectrode, the continuous, high-efficient and high-throughput fusion of the cells in the device is realized.

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. The patent of the invention relates to the precise control of cells in cell electrofusion and the high-efficiency fusion of cells, and is applicable to genetics, animal and plant distant hybridization 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. [0003] Cell electrofusion can be divided into two stages: cell q...

AI Technical Summary

Problems solved by technology

[0007] However, 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 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.

[0010] However, there are still some 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. capable of electrofusion

However, Zhao Zhiqiang proposed that the number of microelectrodes integrated on the chip is small, and high-throughput fusion cannot be achieved; the electric field strength and electric field gradient generated by the microelectrodes are relatively weak, which is relatively weak in terms of precise control of cells; in terms of the selection of processing materials, corrosion resistance, The anti-oxidation ability is also weak; in addition, because the chip uses a planar two-dimensional microelectrode array, there is a great risk of clogging when the cells flow inside the chip; at the same time, because the sample inlet and outlet device is not integrated, the operation is also inconvenient

The method proposed by Japanese researchers is inefficient, and the fusion flux is far from enough to meet the research requirements of fused cells

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