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Cell electrofusion chip device based on micro-chamber array structure

An array structure and array chip technology, which is applied in the field of providing cell electrofusion chips and biological cell electrofusion devices, can solve the problems of large chip spacing, increased system cost, and difficult precise control of cells, so as to reduce requirements and queue up The effect of high rate and high pairwise fusion rate

Inactive Publication Date: 2012-04-18
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
Comparison scheme
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 , and a metal micro-electrode array 4 , a polymer side wall 7 and a polymer water fence 6 are formed on the quartz base layer 8 by micromachining technology. The metal microelectrode array 4 integrates a large number of metal microelectrodes 5, and two opposite metal microelectrodes 5 and two opposite polymer side walls 7 form a micro chamber 9, and the micro chambers 9 are also arranged in an array. The metal micro-electrode array 4 can be made of gold, platinum and other materials, and the polymer material can be selected from polyimide, parylene and other materials; the depth of the micro chamber 9 is 15 μm, the width is 5-3...

Embodiment 2

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

[0052] 1. The micro-chamber array chip is realized by MEMS processing technology. 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. Select quartz as the substrate for processing chips;

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

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

[0056] D. The shape of the micro-electrode array is etched on the Ti / W / Au layer by 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 (15μm);

[0058] F. Photolithography of polyimide to form sidewall structures of tiny cha...

Embodiment 3

[0065] Example 3: Application of Cell Electrofusion

[0066] From the injection port 14 described in Example 1, the cell suspension is injected using a micropump; when the cell suspension enters the storage tank 12 through the passage formed by the conduit 10, the injection port 14 and the microchannel 13, the cells will Under the action of gravity, it will sink into the lower micro chamber 9 corresponding to the sample storage tank 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 path control device 2. Use a micro pump to inject the cell electrofusion buffer; because the micro chamber 9 is a micro pit structure, under the blocking of the polymer side wall 7, the two cells in the micro chamber 9 will not be flushed out by the micro flow; and The excess cells stacked on it will flow out along the microchannel 13, the sample outlet 15 and the conduit 1...

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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 boardto 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 present invention relates to a device for electrofusion of biological cells. Specifically, the present invention relates to a chip that provides cell electrofusion, providing and generating the electric field strengths and electric field gradients required for cell queuing, electroporation, and fusion. The invention relates to precise control of cells and efficient fusion of cells in cell electrofusion, and is suitable for fields such as genetics, animal and plant distant hybrid breeding, developmental biology, drug screening, monoclonal antibody preparation, and mammalian cloning. 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, and suitability for instrument application and standardized operation. [0003] Cell electrofusion can be divided into two main stages: cell queuing and cell fus...

Claims

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Application Information

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Patent Type & Authority Patents(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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