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Cell electrofusion chip device based on micro-chamber array structure

A technology of array structure and array chip, which is applied to the device of biological cell electrofusion and provides the field of cell electrofusion chip, which can solve the problems of precise control of difficult cells, large chip spacing, and increased system cost, and achieve a high fusion rate of two pairs , lower requirements and higher queuing rate

Inactive Publication Date: 2010-04-14
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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.
[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 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
The method proposed by Japanese researchers is inefficient, and the fusion throughput is far from meeting the requirements of post-fusion cell research

Method used

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  • Cell electrofusion chip device based on micro-chamber array structure
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  • Cell electrofusion chip device based on micro-chamber array structure

Examples

Experimental program
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Effect test

Embodiment 1

[0046] see figure 1 , figure 2 , the cell electrofusion microelectrode array chip device based on the micro-chamber structure is composed of a micro-chamber array chip 1 , a flow path control module 2 and a peripheral printed circuit board 3 .

[0047] see figure 2 , image 3 and Figure 4 The micro-chamber array chip 1 has a quartz base layer 8 on which a metal micro-electrode array 4 , polymer side walls 7 and polymer water barriers 6 are formed using micromachining techniques. The metal microelectrode array 4 integrates a large number of metal microelectrodes 5 , and two opposite metal microelectrodes 5 and two opposite polymer sidewalls 7 constitute a tiny chamber 9 , and the microchambers 9 are also arranged in an array. Materials such as metal microelectrode array 4 can be selected from gold and platinum, and polymer materials can be selected from materials such as polyimide and parylene; the depth of the micro-chamber 9 is 15 μm, the width is 5-30 μm, and the leng...

Embodiment 2

[0051] Embodiment 2: the processing technology of part structure in the above-mentioned device

[0052] 1. The micro-chamber array chip is realized by MEMS processing technology, and the processing steps are as follows (the metal array electrode layer material described here is gold Au, and the polymer layer material is polyimide):

[0053] A. Use quartz as the substrate for processing chips;

[0054] B. sputtering a layer of Ti / W on the quartz substrate, wherein the thickness of the Ti / W layer is 50nm;

[0055] C. Electroplate a layer of gold on the surface of the Ti / W layer by electroplating, with a thickness of 15 μm;

[0056] D. Etch the shape of the microelectrode array on the Ti / W / Au layer by means of photolithography;

[0057] E. Spin-coat polyimide glue on the surface of the above structure to form a polyimide layer with a thickness equivalent to the thickness of the Ti / W / Au layer (15 μm);

[0058] F. Photolithography of polyimide to form the sidewall structure of t...

Embodiment 3

[0065] Example 3: Application of Cell Electrofusion

[0066] From the sample inlet 14 described in embodiment 1, use micropump to inject cell suspension; Under the action of gravity, it will sink into the lower micro chamber 9 corresponding to the sample reservoir 12; the special structure of the micro chamber 9 can only accommodate 2 cells in a single layer, and the excess cells will accumulate on it; at this time, combined with the flow control device 2. Use a micropump to inject the cell electrofusion buffer; since the microchamber 9 is a micropit structure, under the obstruction of the polymer side wall 7, the two cells in the microchamber 9 will not be washed out by the microflow; and The superfluous cells accumulated on it will flow out along the microchannel 13, the sample outlet 15 and the conduit 10 due to the lack of blocking of the micro 7; cells; and then apply a sine wave electrical stimulation signal to the metal microelectrode array 4 through the peripheral pri...

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Abstract

The invention provides a cell electrofusion microelectrode array chip device based on a micro-chamber structure, which consists of a micro-chamber array chip, a periphery printed circuit board and a flow passage control module. The micro-chamber array chip consists of a quartz substrate layer, a metal microelectrode array, polymer side walls and a polymer water surrounding fender, wherein two opposite metal microelectrodes and two opposite polymer sidewalls form one micro-chamber which is arranged in an array shape; the micro-chamber array chip is bonded on the periphery printed circuit board to form electrical connection; the flow passage control module covers the micro-chamber array chip. In the invention, a certain electrical field is formed in the micro-chamber by outside electrical signals so as to control high-efficient queue and electrofusion of cells inside the micro-chamber, thus realizing that only one pair of cells inside the micro-chamber are subjected to fusion, and improving throughout and safety of cell fusion.

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 invention relates to precise control of cells and efficient fusion of cells in cell electric fusion, and is applicable to the fields of genetics, animal and plant distant hybrid breeding, developmental biology, drug screening, monoclonal antibody preparation, mammalian cloning and the like. 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 fu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/42C12N15/02
CPCC12M35/02
Inventor 胡宁杨军郑小林胡南夏斌蒋凤赵丽苹刘琳琳田浩张小玲
Owner CHONGQING UNIV
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