Microbial co-culture method, microfluidic chip and use of microfluidic chip

A microfluidic chip and co-cultivation technology, applied in biochemical equipment and methods, microorganisms, microorganisms, etc., can solve problems such as the inability to simulate the interaction process well, the inability to separate two microorganisms separately, and the uncontrollable cultivation process.

Inactive Publication Date: 2019-06-28
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, most of the existing studies on the interaction between probiotics and pathogenic bacteria are based on changing the conditions of the culture environment of a single microorganism, which makes it impossible to simulate the complex interaction process between two microorganisms in the real environment.
A small number of studies on the interaction between probiotics and pathogenic bacteria by co-cultivation are achieved by direct blending, but the cultivation process of this method is uncontrollable, and the two microorganisms cannot be separated separately after the cultivation, which is not conducive to the follow-up research. carry out
There are also very few special devices, such as a U-shaped tube with a semi-permeable membrane in the middle. Although the two kinds of microorganisms can be separated by physical barriers, the cultivation process is not easy to control and difficult to process.

Method used

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  • Microbial co-culture method, microfluidic chip and use of microfluidic chip
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  • Microbial co-culture method, microfluidic chip and use of microfluidic chip

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preparation example Construction

[0082] In a preferred embodiment, the preparation method of hydrogel microspheres embedding different microorganisms independently comprises the following steps:

[0083] In the microfluidic chip, the microorganisms are suspended in the water phase, and the water phase and the oil phase are mixed to form water-in-oil micro-droplets. Hydrogel microspheres of the microorganisms.

[0084] The emulsification / internal gelation process is carried out on the microfluidic chip, the microorganisms are suspended in the water phase, and the water phase with the microorganisms forms water-in-oil micro-droplets in the oil phase, which is then initiated by the added gel initiator The internal gelation reaction forms hydrogel microspheres that entrap microorganisms.

[0085] Preferably, the water phase is a polymer solution dispersed with insoluble calcium salts.

[0086] A typical insoluble calcium salt is calcium carbonate.

[0087] Preferably, the concentration of the polymer solution is...

Embodiment 1

[0153] A kind of co-culture method of probiotics-pathogenic bacteria, probiotics is bacillus rhamnosus; pathogenic bacteria is pseudomonas aeruginosa; comprises the following steps:

[0154] (a) Prepare hydrogel microspheres for embedding probiotics: add sodium alginate solution, calcium carbonate and probiotics to the aqueous phase injection hole of the probiotic microsphere preparation module; Liquid paraffin and Tween 80 are added to the injection hole of the phase, and glacial acetic acid is added, the water phase and the oil phase are mixed to form water-in-oil micro-droplets, and the micro-droplets trigger internal gelation reaction under the action of the gel initiator , forming hydrogel microspheres embedded with probiotics;

[0155] Among them, the concentration of sodium alginate solution is 0.1g / L; the concentration of calcium carbonate in polymer solution is 1g / L; the inoculation density of probiotics is 10 6 CFU / mL;

[0156] The addition of Tween 80 is 1% (v / v);...

Embodiment 2

[0164] A kind of co-cultivation method of probiotics-pathogenic bacteria, probiotics is Bifidobacterium lactis; Pathogenic bacteria is Staphylococcus aureus; It comprises the following steps:

[0165] (a) Prepare hydrogel microspheres for embedding probiotics: add pectin solution, calcium carbonate and probiotics to the water phase injection hole of the probiotic microsphere preparation module; Liquid paraffin and Tween 20 are added to the injection hole, and glacial acetic acid is added, the water phase and the oil phase are mixed to form water-in-oil micro-droplets, and the micro-droplets trigger internal gelation reaction under the action of the gel initiator, Formation of hydrogel microspheres entrapped with probiotics;

[0166] Wherein, the concentration of pectin solution is 5g / L; The concentration of calcium carbonate in polymer solution is 50g / L; The inoculation density of probiotics is 10 8 CFU / mL;

[0167] The addition of Tween 20 is 5% (v / v); The addition of glaci...

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Abstract

The invention discloses a microbial co-culture method, a microfluidic chip and use of the microfluidic chip, and relates to the technical field of biological culture. The microbial co-culture method comprises the following steps that hydrogel microspheres embedding different microorganisms are prepared by using the microfluidic chip separately and conveyed to the same area of the microfluidic chipfor co-culture, wherein the varieties of the microorganisms is no less than two. The microbial co-culture method achieves co-culture of a plurality of varieties of microorganisms by a microfluidic chip technology and a hydrogel microsphere encapsulation technology, and can well simulate a complicated interaction process between two or more varieties of microorganisms in a real environment; separate separation of the later microbial culture is easy, the co-culture and separate separation of the microorganisms are achieved, technical parameters of the microfluidic chip can be controlled, and the research of the co-culture process is facilitated.

Description

technical field [0001] The invention relates to the technical field of biological culture, in particular to a microbial co-cultivation method, a microfluidic chip and its application. Background technique [0002] The discovery and use of antibiotics is one of the greatest contributions to the field of medicine and health in the 20th century. It has an outstanding role in reducing human morbidity and mortality caused by bacterial or viral infections. However, the abuse of antibiotics has also brought new crises to mankind. According to statistics, by 2002, the amount of antibiotics used each year reached 100,000-200,000 tons. Since the 1940s, the total output of antibiotics has exceeded 1 million tons, which has largely contributed to the emergence of a large number of multidrug-resistant "super bacteria", declaring that human beings have officially entered the "post-antibiotic" era. [0003] The treatment of multidrug-resistant pathogenic bacteria infection is a scientific...

Claims

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Application Information

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IPC IPC(8): C12N1/00C12N11/10C12N11/08C12N11/04C12M1/00
Inventor黄术强高梦傅雄飞刘陈立于跃温慧朱伟
OwnerSHENZHEN INST OF ADVANCED TECH