Surface-electrode-technology-based high-throughput cell electrofusion chip device

A surface electrode, high-throughput technology, applied in the field of cell electrofusion, can solve problems such as cell clogging

Inactive Publication Date: 2011-09-07
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0022] The purpose of the present invention is to address the deficiencies in the prior art and propose a high-throughput cell electrofusion chip device based on surface electrodes, which integrates an arrayed microelectrode structure on the chip

Method used

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  • Surface-electrode-technology-based high-throughput cell electrofusion chip device
  • Surface-electrode-technology-based high-throughput cell electrofusion chip device
  • Surface-electrode-technology-based high-throughput cell electrofusion chip device

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0040] Example 1

[0041] See figure 1 , figure 2 with image 3 , A high-throughput cell electrofusion chip device based on surface electrode technology is composed of a high-throughput cell electrofusion chip 1 and a cover sheet 2 with inlet and outlet catheters.

[0042] See image 3 , The high-throughput cell electrofusion chip 1 includes a quartz substrate layer 3, a polymer microchannel layer 5, and a top protective layer 8 from bottom to top.

[0043] The bottom comb-shaped microelectrode array 4, the sidewall surface electrode layer 6, the top comb-shaped microelectrode array layer 7, which are characterized by the bottom comb-shaped microelectrode array layer 4, the sidewall electrode layer 6, and the top comb-shaped microelectrode array layer 7 It is electrically connected to form a "sandwich" structure of the side wall surface electrode array 9, which combines the quartz substrate layer 3 and the polymer microchannel layer 5 to form a smooth flow path 10.

[0044] The bott...

Example Embodiment

[0051] Example 2: How to use this device

[0052] In the experiment, a micro pump can be used to inject the cell suspension into the chip from the injection port 18 in Example 1. When the cell suspension flows through the microchannel 10, the sine wave electrical stimulation signal is loaded through the bonding point 13 or 15. A non-uniform gradient electric field will be formed between the opposite sidewall microelectrodes, and the two cells between the microelectrodes will queue up under the action of the dielectrophoresis force. After the queue is completed, a square wave pulse sequence signal is applied, and the cell pairs that have completed the queue will complete the cell electroporation-cell electrofusion process under the action of the high-intensity pulse electric field between the microelectrodes. After the cell electrofusion process is completed, the micro pump is used to inject the cell culture medium, and the cells in the microchannel flow out through the microchann...

Example Embodiment

[0053] Example 3: Fabrication of high-throughput cell electrofusion chip device

[0054] Using MEMS processing technology, the processing technology is as follows:

[0055] A. Clean the quartz glass sheet;

[0056] B. Place the quartz glass sheet in an evaporation furnace, and evaporate and deposit 2000 的金膜;

[0057] C. Spin-coating AZ4620 on the surface of the gold film, and after exposure and development, it forms the same pattern as the bottom microelectrode array structure;

[0058] D. Put the quartz glass sheet in the gold etching solution and HF etching solution successively to remove the excess gold film to form the bottom microelectrode array structure;

[0059] E. Acetone removes the residual AZ4620 on the surface of the bottom microelectrode array structure;

[0060] F. Cleaning;

[0061] G. Spin-coating PI7510 on the surface of the quartz glass sheet, exposing and developing to form a polymer microchannel layer, and then placing it in a curing oven to solidify PI7510 to form a ...

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Abstract

The invention relates to a surface-electrode-technology-based high-throughput cell electrofusion chip device. The device consists of a microelectrode array chip and a cover plate with a sample charging and discharging guide pipe, wherein the microelectrode array chip comprises a quartz basal layer, a polymer microchannel layer and a top protective layer from bottom to top in turn; a bottom comb-shaped microelectrode array is manufactured on the quartz basal layer; the middle of the polymer microchannel layer is provided with a microchannel; a top comb-shaped microelectrode array of which the shape and the position correspond to those of the bottom comb-shaped microelectrode array is manufactured on the polymer microchannel layer; the side wall of the microchannel is provided with a side wall surface electrode; and the side wall surface electrode is connected with each pair of comb teeth corresponding to the bottom comb-shaped microelectrode array and the top comb-shaped microelectrodearray, so that the side wall surface and the top and bottom comb-shaped microelectrode arrays are electrically communicated to form a sandwich structure. The microelectrode arrays are attached to thepolymer microchannel layer, so that the problem of cell blockage caused by the conventional toothed protruding electrode structure is solved, and the integration level and high fusion efficiency of microelectrodes are ensured simultaneously.

Description

technical field [0001] The present invention relates to cell electrofusion technology, in particular to a high-throughput cell electrofusion chip device based on surface electrodes. In the research of transfection, 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. [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 a non-uniform electric field, they are polarized by the electric field to form a dipole, and the dipole will move under the action of a specif...

Claims

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Application Information

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IPC IPC(8): C12M1/42C12N15/02
CPCC12M35/02
Inventor 胡宁杨军郑小林罗洪艳廖彦剑侯文生许莎张小玲刘琳琳胡晋豪徐涛
Owner CHONGQING UNIV
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