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Microfluidic chip allowing different cell-density arrangement and use thereof

A microfluidic chip and cell technology, applied in the field of combining microstructure technology and microbiology devices

Inactive Publication Date: 2012-11-28
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this device requires manual replacement of cell solutions of different densities to obtain cell colonies of different densities

Method used

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  • Microfluidic chip allowing different cell-density arrangement and use thereof
  • Microfluidic chip allowing different cell-density arrangement and use thereof
  • Microfluidic chip allowing different cell-density arrangement and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Preparation of microfluidic chip

[0032] 1) Draw with L-Edit, make such as figure 1 Graphics, printed on plastic film or optical mask, as exposure mask. The design includes a three-layer structure, which are the cell arrangement chamber 3, the connecting channel 4, the liquid absorption chamber 1, and the fence channel 2 connecting the cell arrangement chamber 3 and the liquid absorption chamber 1 but blocking the passage of cells. The liquid absorption chamber 1, the cell arrangement chamber 3 and the connecting channel 4 are printed on the plastic film, and the fence channel 2 is printed on the optical mask.

[0033] 2) Use the mask pattern obtained in step 1) to prepare a photoresist mold on the silicon wafer through multi-layer alignment sleeve, in which the layer height of the cell arrangement chamber is 20 μm, and the height of the liquid absorption chamber layer is 70 μm. The height of the barrier channel layer is 5 μm. After the overlay is complete...

Embodiment 2

[0036] A microfluidic chip was prepared by the same steps as in Example 1, and each chip included 100 independent cell density arrangement units. Cell arrangement chamber area of ​​200 × 200 μm constant for all independent units 2 A square whose height is 10 μm, while the area of ​​the liquid-absorbing chamber contains 200 × 200 μm 2 , 200×400μm 2 , 200×600μm 2 , 200×800μm 2In four cases, the height is 20 μm. The barrier channel consists of 20 thin channels with a length of 200 μm, a width of 5 μm, and a height of 10 μm.

[0037] After the mold is prepared, use 3mm thick PDMS (A:B=10:1) to replicate the pattern, cure at 75°C for 30 minutes, cut out the patterned PDMS, and naturally adhere to the petri dish. The chip was irradiated under ultraviolet light for 12h, ready for use.

Embodiment 3

[0039] A microfluidic chip was prepared by the same steps as in Example 1, and each chip included 20 independent cell density arrangement units. The cell arrangement chamber area of ​​each unit includes 100×200μm 2 , 200×200μm 2 , 200×300μm 2 There are 5 chambers for cells of different areas arranged in a square and a rectangle. The area of ​​the liquid absorption chamber is 200×200 μm 2 , the height of which is 20 μm. The barrier channel consists of 20 thin channels with a length of 200 μm, a width of 10 μm, and a height of 5 μm.

[0040] After the mold is prepared, use 6mm thick PDMS (A:B=10:1) to replicate the pattern, cure at 80°C for 30 minutes, cut out the patterned PDMS, and naturally adhere to the petri dish. The chip was irradiated under ultraviolet light for 10 h, ready for use.

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Abstract

The invention provides a microfluidic chip allowing different cell-density arrangement and a use thereof. The microfluidic chip comprises multiple cell-density arrangement units. Each one of the multiple cell-density arrangement units comprises a connection channel, a cell arrangement chamber, a liquid absorption chamber and a barrier channel, wherein the connection channel is connected to the cell arrangement chamber and the barrier channel is connected to the cell arrangement chamber and the liquid absorption chamber. The microfluidic chip provided by the invention can realize that cell solutions having the same density can be added into a system and cell communities which are automatically arranged in the cell arrangement chamber and have different density values comprise a single cell, several cells or dozens of cells. Cells are located in the relatively enclosed chambers of the microfluidic chip and thus a flowing liquid located outside the relatively enclosed chambers does not influence the cells. The microfluidic chip can store a large amount of a cell culture solution so that enough nutrients can be supplied to cells and thus the microfluidic chip is suitable for long-term culture and observation.

Description

technical field [0001] The invention belongs to the field of combination of microstructure technology and microbiology device, and specifically relates to a microfluidic chip capable of arranging different cell densities and its application. Background technique [0002] The realization of various physiological functions of cells is regulated by many factors in the surrounding microenvironment, such as nutrition, growth factors, gas molecules, and intercellular interactions, etc.; and the strength of the regulatory factors in cell communities with different densities is different. . In order to better study the effects of regulatory factors around cells on cells and cell-cell interactions, it is of great significance to effectively enrich a certain density of cell communities. In the past, people mostly achieved the enrichment of cells of different densities by sowing cells of different densities on culture dishes, which not only consumes more reagents, is time-consuming an...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12N5/00
Inventor 罗春雄杨薇李昭君康显阶
Owner PEKING UNIV
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