Automatic high-throughput single cell capture method based on liquid droplet microfluidic chip

A microfluidic chip, single-cell technology, applied in the intersection of microfluidic technology and cell biology, can solve the problems of cell damage, high cost, inconvenience of wide application, etc. wide range of effects

Inactive Publication Date: 2018-06-05
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, due to the complex structure and high cost of external electrical components, magnetic control components, optical sensors and mechanical controllers, it is inconvenient to be widely used
The use of physical barriers to capture single cells is simple and low-cost, but it is easy to cause cell damage and is not convenient to load dynamic biochemical signal stimulation

Method used

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  • Automatic high-throughput single cell capture method based on liquid droplet microfluidic chip
  • Automatic high-throughput single cell capture method based on liquid droplet microfluidic chip
  • Automatic high-throughput single cell capture method based on liquid droplet microfluidic chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A method for preparing a high-throughput microfluidic chip that automatically captures single cells is as follows:

[0040] (1) Preparation of SU-8 template for microfluidic chip with high-throughput automatic capture of single cells

[0041] The microfluidic chip was prepared by photolithography and etching methods to prepare the SU-8 template with channel part protrusions; first, the thickness of the first layer of SU-8 glue was 100 μm, pre-baked at 95°C for 20 minutes, and the temperature was naturally lowered, and the mask was placed on the SU-8 template. -8 glue plate, UV exposure for 30s, 95°C post-baking for 20min, and natural cooling; secondly, based on the foundation of the first layer of SU-8 glue, the thickness of the second layer of SU-8 glue is 150μm, and 95°C for 30min before baking. Cool down naturally, put the mask on the SU-8 glue plate, expose to ultraviolet light for 40s, bake at 95°C for 40min, and cool down naturally; finally, use ethyl lactate to d...

Embodiment 2

[0045] The density of single cell suspension is 5×10 4 cells / mL, the chip automatically captures single-cell experiments

[0046] The microfluidic chip prepared above was soaked in 75% ethanol, sterilized by ultraviolet irradiation for 1 h, the outlet of the liquid flow channel was blocked, and placed in a vacuum incubator to evacuate for 10 min. breast cancer cells (MCF-7) single cell suspension in 5×10 4 The cell density of cells / mL is added to the chip inlet. At this time, the flow velocity of the single-cell suspension is 0.027m / s. When all the droplet generation units are filled with the single-cell suspension, the outlet of the liquid flow channel is opened, and the The excess liquid in the medium was drained, and after the cells adhered to the bottom surface of the chip, fresh high-glucose DMEM medium was added and cultured in a 37°C incubator. The distribution of single cells in the chip is shown in Figure 4 shown. The cell density was 5 x 10 4 When the cells / mL ...

Embodiment 3

[0048] The density of single cell suspension is 5×10 5 cells / mL, the chip automatically captures single-cell experiments

[0049] The microfluidic chip prepared above was soaked in 75% ethanol, sterilized by ultraviolet irradiation for 1 h, the outlet of the liquid flow channel was blocked, and placed in a vacuum incubator to evacuate for 10 min. breast cancer cells (MCF-7) single cell suspension in 5×10 5 The cell density of cells / mL is added to the chip inlet. At this time, the flow velocity of the single-cell suspension is 0.027m / s. When all the droplet generation units are filled with the single-cell suspension, the outlet of the liquid flow channel is opened, and the The excess liquid in the medium was drained, and after the cells adhered to the bottom surface of the chip, fresh high-glucose DMEM medium was added and cultured in a 37°C incubator. The distribution of single cells in the chip is shown in Figure 5 shown. The cell density was 5 x 10 5 When the cells / mL ...

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Abstract

The present invention provides an automatic high-throughput single cell capture method based on a liquid droplet microfluidic chip, wherein the microfluidic chip comprises two layers, the upper layeris a flow path inlet and outlet layer, the lower layer is a flow path control layer, the flow path inlet and outlet layer has a liquid flow path channel inlet and a liquid flow path channel outlet, and the flow path control layer comprises a single cell capture flow path channel, gas path channels and liquid droplet generation units. According to the present invention, by introducing the gas pressure controllable gas flow path channel, the negative pressure flow path channel can be formed and can automatically suck the single cell suspension into the capture trap so as to conveniently observeand detect the proliferation, the differentiation, the drug reaction and other behaviors of the single cells; the automatic high-throughput single cell capture is achieved by using the liquid dropletmicrofluidic technology and the fluid mechanics principle; and compared to the traditional single cell capture method, the method of the present invention has advantages of simple, convenient and flexible operation, high throughput, no pollution, wide application range, strong scalability and the like.

Description

technical field [0001] The invention belongs to the intersecting field of microfluidic technology and cell biology, and in particular relates to a method for automatically capturing single cells with high throughput based on a droplet microfluidic chip. Background technique [0002] The microfluidic chip laboratory devoted itself to the research of chip electrophoresis in the early stage, and proposed the concept of micro-total analysis system (μ-TAS), and the research work mainly focused on the continuous flow microfluidic system. In recent years, a new branch has emerged in the field of microfluidic chips - discontinuous flow microfluidic systems, namely droplet microfluidic systems. Droplet microfluidic systems use immiscible two-phase fluids to form droplets at the interface of micropores, and the volume of such droplets is usually in the range of nanoliters to picoliters (10 -9 ~10 -12 L) range. Compared with continuous flow systems, droplets have the characteristics...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12N5/00
CPCC12M23/16C12N5/00
Inventor 秦建华张晓庆姜雷苏文涛
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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