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Device and method for operating multiple adherent cells on same substrate

A substrate and cell technology, applied in tissue cell/virus culture devices, biochemical equipment and methods, biochemical instruments, etc., can solve the problems of reducing the stability of the method, difficult to realize, unstable properties of block copolymers, etc.

Inactive Publication Date: 2010-01-13
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Application Information

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Problems solved by technology

The group applied for a patent at the same time, and the application number was 200710098280.4 (its prior application number was 200610089251.7, and the prior application date was 2006.08.11), which constructed a device and adhesion method for adhering various cells to the same substrate; However, its disadvantage is that it requires a long electrochemical desorption process to orderly adhere different kinds of cells on the same surface, which is not conducive to the promotion of this method.
[0005] The applicant's application number is 200710117815.8, and the invention patent application dated June 25, 2007 has constructed a device and an adhesion method for adhering various cells to the same substrate in an orderly array. The process of this method is simple. However, it is impossible to construct ordered multicellular arrays with continuous distance changes
[0006] The applicant's application number is 200710119999.1 (application date August 6, 2007) and 200710119997.2 (application date August 6, 2007) for two invention patent applications to construct an ordered multicellular arrangement with continuous distance changes, and use electric Chemical desorption method and properties of block copolymers of poly(L-lysine)-polyethylene glycol-poly(L-lysine) enable simultaneous binding, release and selectivity of multiple cells on the same substrate release, but the above two methods need to hydrophilize the surface of the polydimethylsiloxane seal during device preparation, which is difficult to achieve in general biological laboratories; at the same time, the polydimethylsiloxane used in the selective release (L-lysine)-polyethylene glycol-poly(L-lysine) block copolymer properties are unstable, thereby reducing the stability of the method, so the above-mentioned device and method are unfavorable for the general biological laboratory to promote

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  • Device and method for operating multiple adherent cells on same substrate

Examples

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Embodiment 1

[0073] 1) Evaporate a 5nm titanium adhesion layer on the surface of a clean glass slide (thickness 0.15mm), and then vapor-deposit a 30nm thick gold layer on it to obtain a substrate required for an experiment;

[0074] 2) Cut the base of step 1) into 2×2cm 2 small pieces, spare;

[0075] 3) Use photolithography to prepare at least one group of convex linear micro-units on a silicon wafer. First, use the graphics software L-edit to design six side-by-side convex strips of the desired pattern. Among them, the three convex strips on the right The strip is a convex strip of equal cross-section, the middle sections of the three convex strips on the right side are parallel to each other, and the two end sections other than the middle section of the first convex strip on the right are perpendicular to the two ends of the middle section of the convex strip. Fold to the right 0.7cm, and then vertically fold to the middle 0.4cm; the two ends of the second convex strip on the right sid...

Embodiment 2

[0088]1) vapor-deposit 10nm of chromium on the surface of a clean glass slide (thickness 0.12mm) as an adhesion layer, and then vapor-deposit a 50nm-thick gold layer on it to obtain a substrate required for an experiment;

[0089] 2) Cut the base of step 1) into 2×2cm 2 small pieces, spare;

[0090] 3) Prepare at least one group of convex line microstructure units on the silicon wafer using photolithography technology, first use the graphics software L-edit to design six side-by-side convex lines of the desired pattern, the six side-by-side convex lines include : There are a total of six convex lines from left to right; the three convex lines on the right are parallel in the middle, and the first convex line on the right is folded vertically to the right by 0.5cm at both ends of the parallel section, and then folded vertically 0.3cm to the middle; the second convex line on the right is vertically folded to the right at both ends of the parallel section by 0.8cm; the third con...

Embodiment 3

[0097] 1) Evaporate 2nm of titanium on the surface of a clean glass slide (thickness 0.15mm) as an adhesion layer, and then vapor-deposit a 20nm thick gold layer on it to obtain a substrate required for an experiment;

[0098] 2) Cut the base of step 1) into 2×2cm 2 small pieces, spare;

[0099] 3) Prepare at least one group of convex line microstructure units on the silicon wafer using photolithography technology, first use the graphics software L-edit to design six side-by-side convex lines of the desired pattern, the six side-by-side convex lines include : A total of six convex lines from left to right; the three convex lines on the right are parallel in the middle, and the first convex line on the right is folded vertically to the right by 0.8cm at both ends of the parallel section, and then folded vertically 0.5cm to the middle; the second convex line on the right is vertically folded to the right at both ends of the parallel section by 1.5cm; the third convex line on th...

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Abstract

The invention relates to a device and a method for adhering multiple cells to the same substrate and operating the cells, which divide a gold surface of the substrate into a modifiable region and an unmodifiable region by using a microflow pipeline of PDMS. The method comprises the following steps: introducing different molecular modifications to different pipeline regions to ensure that the substrate is divided into a cell adhesive region and a cell nonadhesive region; inoculating different cells in different adhesive regions, taking away the PDMS, and adhering multiple cells to the surface in sequence; electrochemically desorbing to destroy combination of molecules and gold in the nonadhesive region to ensure that all kinds of cells can move freely; and selectively modifying two sides of one or more cells as the nonadhesive region, separating other cells by using PBS, removing PDMS, and fixing the selected cells to lead other cells to move freely. The method and the device have simple and easy operation, can control fixation, complete motion or partial motion of different cells to achieve the aim of operating multiple cells at will, and can be used for studying interaction among multiple cells and medicinal screening.

Description

technical field [0001] The invention relates to a device and an adhesion method for adhering and manipulating various cells on the same substrate. Background technique [0002] The current in vitro research system based on the observation and manipulation of only one type of cell has been unable to meet the needs of basic cell biology and pathology research. People often need to observe and manipulate two or more types of cells in vitro under conditions that mimic living organisms. By investigating the interaction between them, we can understand the process of life body development, disease generation and development, and further carry out drug screening and intervention. [0003] Now, under conditions close to living organisms, the device that can be used to study the interaction of two kinds of cells, the Transwell Chamber is commercially available. This product uses a perforated film to separate the two types of cells into upper and lower layers for culture. You can only...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12N5/00
Inventor 陈振玲李勇蒋兴宇
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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