Cell chemotaxis analysis chip and device as well as use method and preparation method of cell chemotaxis analysis chip

A technology for analyzing chips and cells, which is applied in the field of biology, can solve the problems of difficult control of fluid flow, difficult experimental operation, etc., and achieve the effect of convenient and fast cell chemotaxis analysis test, convenient research, and simple steps

Inactive Publication Date: 2015-05-20
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recently, microfluidic devices (Erwin Berthier, Jill Surfus, James Verbsky, Anna Huttenlocher and David Beebe, An arrayed high-content chemotaxis assay for patient diagnosis, Integrative Biology, 2010, 2, 630-638.) are also widely used to study animal Chemotaxis of cells, generally speaking, the experimental operation is more difficult, and it is also difficult to control the influence of fluid flow on the chemotaxis behavior itself

Method used

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  • Cell chemotaxis analysis chip and device as well as use method and preparation method of cell chemotaxis analysis chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Step 1, in figure 1 Use a puncher to punch through the middle circular liquid inlet chamber to obtain a millimeter-scale liquid inlet 1a for liquid inlet;

[0063] Step 2, fixing the cell chemotaxis analysis chip on the cell culture device, so that the cell chemotaxis analysis chip is in a vacuum state;

[0064] In this field, PDMS preplastics (siloxanes with two different chemical functional groups, A:B=10:1) are commonly used to make cell chemotaxis analysis chips. The main reason is that they have good light transmittance and Gas permeability and light transmission make it easy for experimenters to observe its internal conditions with a microscope, and gas permeability makes it easy to form a vacuum state to facilitate subsequent negative pressure operations. For example, the cell chemotaxis analysis chip can be put into a vacuum pump as a whole to use its own The air permeability of the cell chemotaxis analysis chip is evacuated; it is preferably 5-100Pa. Moreover...

Embodiment 2

[0072] Example 2: Analysis of the response of the same kind of cells to different chemokine solutions.

[0073] Step 1, human breast cancer cell line MDA-MB-231 cells were cultured with DMEM medium containing 10% serum (fetal bovine serum, FBS) and 1% penicillin-streptomycin (culture medium containing various amino acids and glucose, Gibco ), the 10% and 1% mentioned here are volume fractions, the same below, at 37°C and 5%CO 2 cultured in an incubator. The dissociated cells were treated with 0.25% Trypsin-EDTA (trypsin cell digestion solution) solution for 3-4 minutes, then neutralized by adding DMEM culture medium containing 10% FBS, centrifuged at 3000rpm, and discarded the supernatant. Wash the cells with 2% serum culture solution, centrifuge and resuspend to obtain a suspension containing cells.

[0074] Step 2, take two cell chemotaxis analysis chips, use a vacuum pressure pump (the model can be Millipore-WP6122050, the manufacturer Beijing Mintaiyuan Technology Co., L...

Embodiment 3

[0078] Example 3: Analysis of the response of different cells to the same chemokine solution.

[0079] Step 1, human breast cancer cell line MDA-MB-231 cells and human breast cancer cell line MCF-7 cells were respectively treated with DMEM culture medium containing 10% serum (fetal bovine serum, FBS) and 1% penicillin-streptomycin ( containing various amino acids and glucose, Gibco), at 37 °C and 5% CO 2 cultured in an incubator. The dissociated cells were treated with 0.25% Trypsin-EDTA (trypsin cell digestion solution) solution for 3-4 minutes, then neutralized by adding DMEM culture medium containing 10% FBS, centrifuged at 3000rpm, and discarded the supernatant. The cells were washed with 2% serum culture solution, centrifuged and resuspended to obtain two suspensions containing cells.

[0080] Step 2, take two cell chemotaxis analysis chips, and after evacuating with a vacuum pressure pump for 10 minutes, add the two cell suspensions into the two cell chemotaxis analysi...

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Abstract

The invention relates to the technical field of biology, and in particular relates to a cell chemotaxis analysis chip. The cell chemotaxis analysis chip comprises a solution inlet chamber, a chemotaxis channel, a barrier channel and a solution absorption chamber, wherein the solution inlet chamber is provided with a solution inlet for feeding a cell solution and is used for accommodating the cell solution, one end of the chemotaxis channel is communicated with the liquid inlet chamber, the other end of the chemotaxis channel is communicated with one end of the barrier channel, the inner diameter of the chemotaxis channel is greater than the diameter of each cell contained in the cell solution, the inner diameter of the barrier channel is less than the diameter of each cell contained in the cell solution, the other end of the barrier channel is communicated with the solution absorption chamber, and the solution absorption chamber is used for absorbing the solution in the cell solution. The invention also relates to a use method and a preparation method of the cell chemotaxis analysis chip. According to the cell chemotaxis analysis chip provided by the invention, a cell chemotaxis process at a specific concentration gradient can be quantitatively observed very conveniently, and the influence on the chemotaxis process by flow is eliminated to facilitate the research on the cell chemotaxis behavior.

Description

technical field [0001] The invention relates to the technical field of biology, in particular to a cell chemotaxis analysis chip, device, use method and production method. Background technique [0002] Chemotaxis behavior is a basic life process, and its research is not only beneficial to understanding the response mechanism of biological signal transduction, but also has very high practical value in the development of related biosensors. However, the current research methods are still relatively backward. For the research methods of cell chemotaxis, the current experimental operation is more difficult and inconvenient to use. [0003] Microfluidic technology is a cutting-edge subject that has developed rapidly in recent years. With its advantages of diversification, fast analysis, low dosage, high throughput and real-time observation, it has become a powerful research method in the field of biological cell analysis. Recently, microfluidic devices (Erwin Berthier, Jill Surf...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/34C12Q1/02
Inventor 罗春雄杨薇冉敏欧阳颀
Owner PEKING UNIV
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