A kind of cell in situ electroporation device and using method

Abstract

The invention relates to a cell in-situ electroporation device and a use method. The cell in-situ electroporation device is characterized by comprising a substrate and at least two electrode pairs; each electrode pair comprises a first electrode and a second electrode which are arranged in a crossed mode, and cathodes and anodes of the first electrodes and the second electrodes are arranged oppositely; the electrode pairs are fixedly arranged on the substrate, the first electrodes and the second electrodes of the two adjacent electrode pairs are arranged in a nested mode, the electrode pairs are both electrically connected with an external electric pulse generator, and the electric pulse generator applies electric pulses to the electrode pairs in sequence in a space coding mode; two protruding structures are arranged at the two ends of the bottom of the substrate in an inward extending mode respectively; by means of the protruding structures, the electroporation device is supported above cells at the bottom of a culture dish, and the electrode pairs exactly face the cells for electroporation. The cell in-situ electroporation device can be widely applied to cytobiology experimental research.

Description

technical field [0001] The invention relates to a cell in situ electroporation device and a using method, belonging to the field of electroporation. Background technique [0002] Electroporation is a kind of transmembrane voltage generated by the cell membrane by applying an external pulse electric field. When the cell transmembrane voltage exceeds a certain threshold, a recoverable micropore is formed on the surface of the cell membrane to introduce foreign substances into the cell. A cell transfection or bacteria method of conversion. Electroporation has the advantages of non-cytotoxicity, low cost, and high transfection efficiency. At present, electroporation has been widely used in biological research. Most of the existing commercial electroporation instruments use parallel electrodes to perforate between electrodes. Cell solutions, such as placing cells between parallel electrodes a few millimeters apart or allowing cells to flow between parallel electrodes, but there ...

AI Technical Summary

Problems solved by technology

Electroporation has the advantages of non-cytotoxicity, low cost, and high transfection efficiency. At present, electroporation has been widely used in biological research. Most of the existing commercial electroporation instruments use parallel electrodes to perforate between electrodes. Cell solutions, such as placing cells between parallel electrodes a few millimeters apart or allowing cells to flow between parallel electrodes, but there is a common problem that can only deal with cells in suspension, and if transfected adherent cells Not only is the protocol cumbersome to digest and resuspend cells into electroporation buffer, but digesting cells can also affect the viability of adherent cells, especially for primary cells

[0003] Aiming at the deficiency that adherent cells need to be digested before electroporation in the traditional method, the prior art discloses a microelectrode array chip to transfect adherent cells in situ, arrange finger electrodes on the bottom of the multi-culture dish, and then place the cells Directly cultivated on the bottom of the petri dish, and apply electrical pulses directly through the electrodes at the bottom of the petri dish to transfect the cells above the electrodes and in the electrode gap area, but these electrodes often need to be fabricated using expensive micro-processing techniques, and the cells and electrodes are in direct contact, The cells on and near the electrode will be damaged by the heat generated by electrolyzed water and the change of pH value, and the distribution of the electric field is significantly uneven, which will further affect the effect of cell transfection

[0004] In order to avoid the damage of cells directly touching the electrodes or being very close to the electrodes, Garcia-Sanchez et al. developed a non-contact in situ electroporation device using PCB as electrodes. The electrodes are suspended above the cells and do not directly contact the cells. Avoid the damage caused by water electrolysis during electroporation. However, this method still has the problem of uneven electric field distribution, and cannot achieve efficient and uniform electroporation transfection for adherent cells.

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