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Micro-fluidic chip based on droplet micro-fluidic technology and detection method of micro-fluidic chip

A microfluidic chip and microfluidic technology technology, applied in biochemical equipment and methods, enzymology/microbiology devices, tissue cell/virus culture devices, etc., can solve the problem of low single-cell capture rate, single-cell secretion The problem of low detection sensitivity and inability to directly detect cell secretions in real time can achieve the effect of simple and flexible operation, short time-consuming and low cost.

Active Publication Date: 2021-03-23
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In view of the shortcomings of the prior art described above, the purpose of the present invention is to provide a microfluidic chip based on droplet microfluidic technology and its detection method, which is used to solve the problem of single-cell defects in the microfluidic chip in the prior art. Problems such as low capture rate, low sensitivity of single-cell secretion detection, and inability to directly detect cell secretion in real time

Method used

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  • Micro-fluidic chip based on droplet micro-fluidic technology and detection method of micro-fluidic chip
  • Micro-fluidic chip based on droplet micro-fluidic technology and detection method of micro-fluidic chip
  • Micro-fluidic chip based on droplet micro-fluidic technology and detection method of micro-fluidic chip

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Embodiment 1

[0064] Such as Figure 1 to Figure 4 As shown, this embodiment provides a microfluidic chip based on droplet microfluidic technology, and the microfluidic chip includes: a droplet generation and single cell capture unit 2 located on the upper layer and a single cell culture unit located on the lower layer ;

[0065] Such as figure 1 and figure 2 As shown, the droplet generation and single cell capture unit 2 includes: a continuous phase inlet 10, a first dispersed phase inlet 11, a second dispersed phase inlet 12, a dispersed phase liquid inlet channel 13, a continuous phase liquid inlet channel 14, ten Zigzag droplet generation channel 15, liquid outlet channel 16 and liquid outlet 17;

[0066] Such as figure 1 As shown, the dispersed phase liquid inlet channel 13 includes three: an arc-shaped dispersed phase liquid inlet channel 131, a spiral dispersed phase liquid inlet channel 132 and a linear dispersed phase liquid inlet channel 133. The first dispersed phase inlet 1...

Embodiment 2

[0079] This embodiment provides a method for preparing a microfluidic chip based on the droplet microfluidic technology described in the above-mentioned embodiment 1, comprising the following steps:

[0080] Step S1, using AutoCAD software to draw the channel structure diagram of the droplet generation and single cell capture unit 2 in the microfluidic chip, and making a mask according to the diagram.

[0081] Step S2, using photolithography and etching methods on the 4-inch single crystal silicon wafer, through glue coating, glue removal, pre-baking, exposure, post-baking, development, and hard baking, to prepare a SU-8 template with channel protrusions. Specifically: cast a layer of SU-8 glue on a 4-inch single-crystal silicon wafer to make it thick between 50 μm and 60 μm, pre-bake at 95°C for 40 minutes, and naturally cool to room temperature; place the mask on the single-crystal silicon wafer On the top of the silicon wafer, use a photolithography machine for UV exposure ...

Embodiment 3

[0085] This embodiment provides a detection method for a microfluidic chip based on droplet microfluidics technology, the detection method comprising the following steps:

[0086] Step S1, providing a microfluidic chip based on droplet microfluidic technology as described in Example 1. The microfluidic chip can be prepared by the preparation method described in Example 2, but is not limited thereto, and can also be prepared by other preparation methods.

[0087] Step S2, using a pressure pump to make the aqueous phase single-cell suspension enter from the second dispersed phase inlet 12, the aqueous phase magnetic bead suspension enter from the first dispersed phase inlet 11, and the oil phase liquid enter from the continuous phase inlet 10 into, wherein, the aqueous phase single cell suspension contains basement membrane matrigel.

[0088] As an example, the pressure pump is a multi-channel pressure pump, which can simultaneously apply pressure to the second dispersed phase ...

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Abstract

The invention provides a micro-fluidic chip based on a droplet micro-fluidic technology and a detection method of the micro-fluidic chip. Cells can be monodispersed by adopting a spiral disperse phaseliquid inlet channel according to an inertial focusing principle, so that the cells are uniformly distributed in the channel from front to back, and single-cell droplets with high wrapping rate are favorably formed; besides, the disperse phase liquid inlet channel and a continuous phase liquid inlet channel are intersected in a cross shape to form a cross-shaped droplet generation channel, the flow ratio of two-phase liquid is convenient to adjust, the length and the distance of generated droplets are controlled, and the sizes of the droplets are more uniform and stable; and moreover, the micro-fluidic chip is provided with an upper layer and a lower layer, the upper layer is used for capturing high-flux single cells, and the lower layer is used for culturing the single cells and enriching secreta of the single cells, so that the single cells are cultured in droplets for a long time, and researches such as single cell in-situ culture, cell co-culture, drug screening, real-time and high-sensitivity detection of the secreta can be carried out, and the micro-fluidic chip is simple and flexible to operate, high in flux, pollution-free, short in consumed time, low in cost and wide in application range.

Description

technical field [0001] The invention belongs to the technical field of cell culture and biological detection, and in particular relates to a microfluidic chip and a detection method based on droplet microfluidic technology that integrate single cell capture, culture, and real-time detection of secretions. Background technique [0002] Cells are the basic unit of organism structure and function, and in any cell population, there will be different degrees of cell heterogeneity, and cell heterogeneity has important roles in tumor progression, tumor drug resistance, immune response, cell differentiation, etc. Significant influence. In recent years, with the continuous development of biology and medicine, the research on single cells has become more and more important. Through the study of single cells, we can see the influence of cell heterogeneity on the function and state of the matrix, and the study of the pathological characteristics of different single cells is of great si...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M1/22C12M1/00
CPCC12M23/16C12M23/10C12M47/04
Inventor 贾春平刘伟周扬吴嫚赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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