Apparatus for ordered adhering various cells to setting position of same substrate and adhesion method

A technology for setting position and cells, applied in biochemical equipment and methods, immobilized enzymes, microbial measurement/testing, etc., can solve the problem of cell patterns that cannot interact, operate complicatedly, and cannot control adhesion well Sports and other issues

Active Publication Date: 2009-02-11
重庆龙翼生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to overcome the complex operation of the prior art when adhering various cells to the same substrate surface, and the movement of the adhered cells cannot be well controlled, so that the fixed cell pattern is destroyed or cannot interact at all. , so as to provide a device and an adhesion method for orderly adhering various cells to the set position of the same substrate, the device has a simple structure and is easy to operate, and can effectively control the distance between different cells The movement between different kinds of cells is convenient for studying the interaction between different kinds of cells

Method used

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  • Apparatus for ordered adhering various cells to setting position of same substrate and adhesion method
  • Apparatus for ordered adhering various cells to setting position of same substrate and adhesion method
  • Apparatus for ordered adhering various cells to setting position of same substrate and adhesion method

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Experimental program
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Effect test

Embodiment 1

[0078] Please refer to Figure 2-1, Figure 2-2, Figure 2-3 and Figure 2-4:

[0079] 1) Evaporate a 10nm (2-10nm) titanium adhesion layer on the surface of a clean glass slide 1 (thickness 0.15mm), and then vapor-deposit a 40nm (20-50nm) titanium adhesion layer on the surface Thick gold layer, making substrate 1;

[0080] Cut the prepared substrate 1 into 2 x 2cm 2 Soak the small pieces in an aqueous solution of ethanol (concentration: 50% to 100%), then blow dry with nitrogen for subsequent use;

[0081] 2) Use photolithography technology to engrave a pattern with at least one group of convex strip microstructure units on a silicon wafer. The manufacturing steps are as follows: first use the drawing software L-edit to design seven parallel convex strips of the desired pattern. Shaped lines, the seven parallel convex lines form a set of convex strip microstructure units, including:

[0082] Straight-line central convex strip in the middle position;

[0083] The first convex ...

Embodiment 2

[0091] 1) Evaporate a 5nm (2-10nm) chrome adhesion layer on the surface of a clean glass slide (thickness 0.15mm), and then vapor-deposit a 20nm (20-50nm) thickness on the chromium adhesion layer The gold layer is made substrate 1;

[0092] Cut the prepared substrate 1 into 2 x 2cm 2 Soak the small pieces in an aqueous solution of ethanol (concentration: 50% to 100%), then blow dry with nitrogen for subsequent use;

[0093]2) Use photolithography technology to engrave a pattern with at least one group of convex strip microstructure units on a silicon wafer. The manufacturing steps are as follows: first use the drawing software L-edit to design seven parallel convex strips of the desired pattern. Shaped lines, the seven parallel convex lines form a set of convex strip microstructure units, including:

[0094] Straight-line central convex strip in the middle position;

[0095] The first convex strip, the second convex strip and the third convex strip arranged in sequence on o...

Embodiment 3

[0103] The substrate obtained in embodiment 1 or embodiment 2 that has been adhered to various cells is immersed in common cell culture fluid (fetal bovine serum concentration 10%); a positive electrode is connected to the solution, and a negative electrode is connected to the gold surface of the thiolated substrate , the voltage is 1.4V, the power-on time is 30s, and an electrochemical analysis is performed once. The thiol analysis and attachment of the gold surface area outside the specified pattern becomes the surface layer that can adsorb proteins and cells again, and adheres to the specified pattern. The cells are free to move.

[0104] In addition, the convex strips of the convex strip type microstructure unit according to the present invention can also add convex strips with the same shape and parallel to the outermost convex strips as required; the microgrooves The concave grooves of the unit can also be added with the same shape and parallel concave grooves outside th...

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Abstract

The invention relates to a device which conglutinates multiple cells to the set position of the same substrate, a preparation method and the usage thereof; the device comprises the substrate which is plated with a titanium or chromium adhesion layer and a golden layer on the upper surface in a way of evaporation, a poly-dimethylene siloxane stamp which is treated by oxidation, has a hydrophilic surface and is provided with at least one group of micro chase units on the lower surface; a vertical through hole communicated with the corresponding groove is arranged at the end part of the grooves on the stamp; the lower surface of the stamp is stuck on the golden layer of the substrate and is combined with the golden layer in a reversible way to form a closed pipeline; when the closed pipeline is formed in 10min, FN is pumped into the middle, the second and the fifth grooves of the stamp; subsequently, EG6 is pumped into the first, the third, the fourth and the sixth grooves; after 1-3h, extracellular matrix proteins are formed on the golden layers of the middle, the second and the fifth grooves, and inertia monomolecular membrane layers are formed on the golden layers and the surrounding of the first, the third, the fourth and the sixth grooves. The device orderly conglutinates multiple cells to the same substrate in an adjustable way, and can be used for understanding the action of medicine on different cells as well as for drug screening.

Description

technical field [0001] The present invention relates to a device and an adhesion method for orderly adhering various cells to the same substrate, in particular to a device and an adhesion method for adhering various cells to the same substrate in an orderly and controllable manner . Background technique [0002] In the basic research fields of cell biology and tissue biology, as well as the applied research of biomedical engineering and drug detection based on the interaction between cells, it is often necessary to immobilize two or more kinds of cells on the same surface. Recently, some methods of related methods have been reported. [0003] For example in literature 1: Chiu, D.T.; Jeon, N.L.; Huang, S.; Kane, R.S.; Wargo, C.J.; , (6), 2408-2413, the Whitesides experimental group disclosed a method of immobilizing various cells on the same surface using a three-dimensional microfluidic channel, and can form a complex pattern composed of various cells. This method first p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/00C12Q1/04
Inventor 李勇陈振玲蒋兴宇
Owner 重庆龙翼生物科技有限公司
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