Establishment and application of mastocyte-macrophage-coculture-based microfluidic chip

A microfluidic chip, chip technology, applied in tissue cell/virus culture devices, animal cells, vertebrate cells, etc., can solve problems such as risks that cannot be ignored, unfavorable diagnosis and treatment of allergic diseases, etc.

Inactive Publication Date: 2016-05-04
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although in vivo detection is the most convenient and reliable method for doctors or patients, its potential risks cannot be ignored and need to be treated with caution
In my country, the in vitro detection of allergens has not received much attention, and the traditional diagnostic mode is still in use, which is not conducive to the timely diagnosis and treatment of allergic diseases, but also brings additional physical and mental pain to patients

Method used

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  • Establishment and application of mastocyte-macrophage-coculture-based microfluidic chip
  • Establishment and application of mastocyte-macrophage-coculture-based microfluidic chip
  • Establishment and application of mastocyte-macrophage-coculture-based microfluidic chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Microfluidic chip physical map

[0034] The physical picture of the microfluidic chip is as follows figure 1 As shown, the specific operation is as follows:

[0035] (1) The first part is the main structure of the chip - the PDMS flow channel layer, with a total length of 44670.83 μm, a width of 31200 μm, and a thickness of 4 mm; two long DC channels with a width of 1000 μm and a length of 31673.16 μm are designed on the chip, and the two ends are respectively equipped with diameter The entrance and exit holes are 2500μm.

[0036] (2) A capillary channel with a length of 10 mm and a width of 100 μm is connected between the two channels.

[0037] (3) Open holes with a diameter of 2 mm are provided at both ends of the capillary to facilitate the insertion of the miniature reference electrode.

[0038] (4) Four sets of gold electrodes are electroplated and integrated on the bottom surface of the cell chip, and a three-electrode system is formed with a plug-in...

Embodiment 2

[0040] Example 2 Dimensions of the gold-plated layer of the microfluidic chip

[0041] Such as figure 2 As shown, the specific operation is as follows:

[0042] (1) Electroplate four groups of gold electrodes on its surface, and the diameter of each working electrode is 1000 μm.

[0043] (2) The guide wire with a diameter of 500 μm connects the working electrode and the gold finger (fixed by the electrode clamp) with a width of 4000 μm on the edge of the chip. In front of the working electrode is an electroplated gold layer (counter electrode) with a half-circle width of 1000 μm. It is also connected to the gold finger on the edge of the chip by a 500μm wide plating wire.

Embodiment 3

[0044] Example 3 Schematic diagram of microfluidic chip assembly

[0045] The entire microfluidic chip was fabricated using a soft lithography method, using polydimethylsiloxane as the main material, which has the characteristics of good biocompatibility and gas permeability (such as image 3 ). The specific operation is as follows:

[0046] (1) Produce SU-8 negative glue (SU82075) master plate on the cleaned silicon wafer by standard photolithography process, and complete the basic production after pre-baking, mask covering, UV exposure, post-baking and developing the master plate .

[0047] (2) Secondly, the PDMS substrate containing microchannels was made by casting method. After the mold was hardened at 65°C for 5 minutes, the PDMS prepolymer and crosslinking agent (mass ratio: 10:1) After mixing evenly, pour it into a mold and dry it in an oven at 80°C for 2 hours.

[0048] (3) After the polymerization is completed, the PDMS is peeled off from the mold, and holes such...

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Abstract

The invention discloses establishment and application of a mastocyte-macrophage-coculture-based microfluidic chip, belonging to the field of food quality analysis and detection. Special flow channel design and tensile valve channels are adopted to implement non-direct contact coculture on the two cells, thereby accurately controlling the coculture process of the two cells and effectively researching the cell paracrine secretion mechanism. The electrochemical technique is combined with the microfluidic chip, and electrochemical resistance signals are adopted on a PDMS (polydimethylsiloxane) substrate surface electrogilding electrode to monitor physiological activities of cells in the coculture in real time. By adopting the design above, the cell coculture microfluidic chip platform for food allergen identification evaluation studies is established. The two-dimensional cell PDMS microfluidic chip platform is established and successfully applied to food allergen identification evaluation studies.

Description

technical field [0001] The invention relates to the construction and application of a mast cell-macrophage co-cultured microfluidic chip, which belongs to the field of food quality analysis and detection. Background technique [0002] Food allergens refer to antigenic molecules in food that can cause the body's immune response. Aquatic products are the main source of human food-derived protein, and allergies caused by crustacean aquatic products such as shrimp and crab are the most common clinical reactions. . Food allergy is a common allergic disease that is widely prevalent in the world. It has the characteristics of various symptoms, complex symptoms, extensive harm, and difficult to cure completely. It has long been a major problem in the medical field. problem. In recent years, with the increasing incidence of food allergy worldwide, the prevention and detection of food allergy has become an urgent need, but because there is no effective treatment for food allergy, av...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12N5/0787C12N5/0786G01N33/50
CPCC12M23/16C12M23/58C12M29/00C12N5/0642C12N5/0645C12N2502/11C12N2502/1157G01N33/5005
Inventor 孙秀兰蒋栋磊孙嘉笛张银志
Owner JIANGNAN UNIV
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