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Visible Transwell chip and preparation method thereof

A chip and top-level chip technology, applied in biochemical equipment and methods, methods of supporting/immobilizing microorganisms, biochemical instruments, etc., can solve the problems of difficulty in realizing Transwell experiments and limitations of microfluidic chips

Inactive Publication Date: 2017-06-09
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

However, microfluidic chips are difficult to realize the functions of Transwell experiments, and are greatly limited in the research of drug metabolism and cell invasion.
[0004] At present, there is no method that can directly observe the phenomenon under the Transwell chamber in real time. It is still in the blank stage to use the visualized Transwell chip to conduct relevant research and analysis. If it can be realized, it will have great application prospects in biological research and pharmaceutical research and development.

Method used

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  • Visible Transwell chip and preparation method thereof
  • Visible Transwell chip and preparation method thereof
  • Visible Transwell chip and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Visual Chip Fabrication

[0025] Since the top chip 1 and the bottom chip 3 are transparent PDMS chips, and the porous filter membrane 2 is opaque, the porous filter membrane 2 is punched at a predetermined position as the observation window 4, so that the porous filter membrane 2 can be observed Window 4 is used to observe the state of cells in channel 6 of the bottom chip. Put the porous filter membrane 2 on a glass sheet for UV activation for 1 hour, then silanize it for 30 minutes, and then place the porous filter membrane 2 and the top chip 1 together for irreversible oxygen plasma sealing, then heat in an oven at 80 degrees for 30 minute. Use PDMS polymer with a ratio of monomer to initiator of 20:1, throw a 10um-50um thick film on a glass plate, treat the upper surface of the bottom chip 3 with oxygen plasma, dip the thin PDMS, and seal it with The porous filter membrane 2 of the top chip 1 is aligned and bonded, heated in an oven at 80 degrees, and cured compl...

Embodiment 2

[0027] Visual Chip Fabrication

[0028] Since the top chip 1 and the bottom chip 3 are transparent PDMS chips, and the porous filter membrane 2 is opaque, in order to observe the cell state in the bottom chip channel 6, the porous filter membrane 2 is cut off at a predetermined position as the observation area 5 . After cutting the porous filter membrane 2 to a suitable size, put it on a glass sheet for UV activation for 1 hour, and then silanize it for 30 minutes, perform oxygen plasma sealing with the top chip 1, and place it in an oven at 80 degrees for 30 minutes. Use PDMS polymer with a ratio of monomer to initiator of 20:1, throw a 10um-50um thick film on a glass plate, treat the upper surface of the bottom chip 3 with oxygen plasma, dip the thin PDMS, and seal it with The porous filter membrane 2 of the top chip 1 is aligned and bonded, heated in an oven at 80 degrees, and cured completely in 30 minutes. After the sealed chip is taken out of the oven, it is cut into t...

Embodiment 3

[0030] Visual chip application

[0031] Put the chip in a petri dish and sterilize it by ultraviolet light for 2 hours. Use a pipette gun to add cell culture medium from the inlets of the bottom chip channel 6 and the top chip channel 7, so that the culture medium fills the entire bottom chip channel 6 and top chip channel 7. Digest the HePG2 cells in the culture flask with trypsin, stop the digestion after the cells fall off the wall, suck the cell suspension in the culture flask into a centrifuge tube for centrifugation, suck the supernatant and throw it away, add Resuspend the cells with a new medium, draw the cell suspension with a pipette gun, add it from the inlet of channel 6 of the bottom chip, suck out the corresponding volume of medium from the outlet of the liquid, and replace it. After repeating three times, the bottom chip can be regarded as Channel 6 has been filled with cell suspension, and medium is added to the culture dish to reduce the volatilization of the ...

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Abstract

The invention provides a visible Transwell chip and a preparation method thereof, and particularly relates to a micro-fluidic chip which carries a porous filter membrane and has a complicated channel structure. The micro-fluidic chip mainly comprises a top chip with a window, a porous filter membrane and a bottom chip, wherein the lower surface of the top chip with a window design is irreversibly sealed by using the porous filter membrane, and the porous filter membrane is connected with the upper surface of the bottom chip through PDMS adhesion. The micro-fluidic chip has functions of a Transwell cell and the micro-fluidic chip, can be used for directly observing the bottom chip through the window, and can be applied to medicine metabolism, cell invasion and other biological researches.

Description

technical field [0001] The invention relates to the field of microfluidic chip preparation, in particular to a visualized Transwell chip and a preparation method thereof. Background technique [0002] Transwell experimental technology, the traditional method is to put the Transwell chamber into the culture plate, the chamber is called the upper chamber, the culture plate is called the lower chamber, the upper chamber is filled with the upper layer culture solution, the lower chamber is filled with the lower layer culture solution, and the upper and lower layer culture solution is made of polycarbonate separated by ester membranes. We planted the cells in the upper chamber, and because the polycarbonate membrane is permeable, the components in the lower layer of the culture medium can affect the cells in the upper chamber, so that we can study the effects of the components in the lower layer of the culture medium on cell growth and movement. However, due to the limitation of...

Claims

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Application Information

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IPC IPC(8): C12M3/06
CPCC12M23/16C12M25/02C12M41/46
Inventor 秦建华尹方超李中玉郭雅琼
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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