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Multifunctional micro-fluidic chip as well as preparation method and application thereof

A microfluidic chip and multi-functional technology, applied in the field of cell biology and microfluidics, can solve the problems of microchannel blockage, premature gelation, uneven distribution of calcium ions, etc., and achieve simplified operation and droplet size Uniformity, Poisson effect reduction effect

Active Publication Date: 2020-06-05
LIAONING UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the actual operation process, conventional 3D droplet generation chips have problems such as uneven distribution of calcium ions, premature gelation, and microchannel blockage.

Method used

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  • Multifunctional micro-fluidic chip as well as preparation method and application thereof
  • Multifunctional micro-fluidic chip as well as preparation method and application thereof
  • Multifunctional micro-fluidic chip as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] A method for making a multifunctional microfluidic chip:

[0069] Using SU-8 negative photoresist technology, multi-layer PDMS thermal bonding technology and plasma bonding technology, and using polydimethylsiloxane (PDMS) with good light transmission, gas permeability and biocompatibility and conventional The glass substrate is used as a material to manufacture an integrated microfluidic chip with a multi-layer structure. The specific steps are as follows:

[0070] 1. Cleaning of monocrystalline silicon wafers: put monocrystalline silicon wafers cut into suitable sizes into piranha solution (concentrated H 2 SO 4 :H 2 o 2 =3:1 volume ratio) soak overnight, take it out, wash it with acetone, absolute ethanol and deionized water respectively, and put it on a heating plate to dry at 120°C for 30min to completely dry the silicon wafer.

[0071] 2. Production of SU-8 silicon wafer positive mold: Pour appropriate amount of SU8-2075 photoresist along the corners of the si...

Embodiment 2

[0077] An application of a multifunctional microfluidic chip, comprising the steps of:

[0078] Instruments and reagents:

[0079] Precision micro-injection pump (LSP04-1A, Baoding Lange Company); Nikon ECLIPSETI inverted fluorescence microscope (Nikon Company, Japan); sodium alginate; calcium chloride; sodium fluorescein; mineral oil; ABIL EM-90; chrysin-7 -O-β-glucuronide, baicalein, oreoside B; DMEM complete medium.

[0080] Preparation of the solution: Weigh appropriate amount of sodium alginate powder and calcium chloride powder and irradiate it under the ultra-clean table ultraviolet lamp for 0.5h, and prepare 1% (w / w) sodium alginate solution and 1% (w / w) sodium alginate solution and 1% ( w / w) calcium chloride solution; take appropriate amount of ABIL EM-90 water-in-oil emulsifier in mineral oil, and finally form 3% ABIL EM-90-mineral oil solution.

[0081] 3D single-cell droplet generation from sodium alginate

[0082] Connect the precision syringe pump to the chip ...

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Abstract

The invention discloses a multifunctional micro-fluidic chip which comprises a micro valve control channel layer, a fluid channel layer and a glass substrate layer which are arranged from top to bottom in sequence, wherein the micro valve control channel layer and the fluid channel layer form a fluid channel unit through PDMS (polydimethyl siloxane) thermal bonding; and the fluid channel unit andthe glass substrate layer are integrally boned through plasmas. Overall three main function structure units of the chip are applied to unicellular liquid droplet generation, cell culture and calcium ion infusion or drug intervention, and a cell culture zone is divided into cell culture tanks of an identical structure. By controlling opening and closure of valves, flexible operation and control ona micro fluid can be achieved; due to adoption of an independent auxiliary fluid infusion unit, problems of gelatinization ahead of time, channel blocking caused by gels, non-uniform distribution of calcium ions, and the like can be avoided; and meanwhile, the stability of liquid droplets is improved through liquid droplet gelatinization. The chip is capable of achieving three functions of unicellular liquid droplet generation, three-dimensional culture and antitumor drug screening simultaneously.

Description

technical field [0001] The invention belongs to the field of cell biology and microfluidic technology, and in particular relates to a microfluidic chip used for single cell droplet generation, three-dimensional culture and antitumor drug screening. Background technique [0002] Drug screening is the initial process and key step in the research of new anti-tumor drugs. However, the research and development of new drugs has disadvantages such as high cost, long cycle, complicated process, and low success rate, which have always restricted the screening and development of modern drugs. At present, drug screening based on 96-well plate is one of the technical means of preliminary screening of drugs widely used by most international drug research institutions. However, this method of drug screening based on conventional orifice plates has some limitations, such as large reagent consumption, cumbersome operation, and low detection sensitivity. In addition, during the operation, th...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12M1/00
CPCC12M23/16C12M33/04
Inventor 孟宪生李慧包永睿王帅李天娇
Owner LIAONING UNIV OF TRADITIONAL CHINESE MEDICINE
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