Micro-fluidic chip for observing and treating suspended cells in real time and preparation method and applications thereof

A microfluidic chip and suspension cell technology, applied in biochemical equipment and methods, biomass post-treatment, biomass pretreatment, etc., can solve the problems of inapplicability and achieve good biocompatibility

Active Publication Date: 2019-04-12
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Suspension cells, another of the two major classifications of cells, req

Method used

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  • Micro-fluidic chip for observing and treating suspended cells in real time and preparation method and applications thereof
  • Micro-fluidic chip for observing and treating suspended cells in real time and preparation method and applications thereof
  • Micro-fluidic chip for observing and treating suspended cells in real time and preparation method and applications thereof

Examples

Experimental program
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preparation example Construction

[0053] The present invention also provides a method for preparing the above-mentioned microfluidic chip for real-time observation and processing of suspended cells, including the following steps:

[0054] The first step is to use computer graphics software to design the chip structure, and make the corresponding photolithography mask through the laser phototypesetting machine;

[0055]In the second step, the photoresist is coated on the surface of the silicon wafer, subjected to ultraviolet exposure and development, and the pattern on the mask plate is transferred to the photoresist to obtain a template of the upper substrate.

[0056] In the third step, the polydimethylsiloxane (PDMS) material is cast on the surface of the template, and the upper substrate is obtained through degassing, baking, and drilling.

[0057] In the fourth step, the polydimethylsiloxane material is coated on the surface of the glass sheet, and baked to obtain the bottom layer.

[0058] The fifth step...

Embodiment 1

[0077] A microfluidic chip for real-time observation and processing of suspended cells, comprising an upper substrate 1 with a microfluidic channel, a PDMS film 2 coated on the surface of the lower substrate, and a lower substrate 3 . The material of the upper substrate 1 of the microfluidic chip and the material of the thin film 2 coated on the lower film are all polydimethylsiloxane materials; the material of the lower film 3 is glass. The microfluidic channel of the upper substrate consists of a cell inoculation channel 13, an oily solution inlet channel 12, an aqueous solution inlet channel 11, a double-emulsion droplet buffer storage area 14, a double-emulsion droplet anchoring area 15, and redundant droplet The discharge channels 16, 17, 18, and the anchor droplet discharge channel 19 constitute. The cell seeding channel 13 is a straight channel with constant width, and the width is 100 μm. The oily solution inlet channel 12 has a width of 100 μm, forming a cross-shaped...

Embodiment 2

[0082] This embodiment is a method for treating the surface hydrophilicity of the microfluidic channel of the microfluidic chip for processing suspended cells described in Example 1, including the following steps:

[0083] In the first step, the aqueous solution of 4wt% polyvinyl alcohol (PVA) is passed into the microfluidic chip from the inlet a of the aqueous solution inlet channel, and the air of 5KPa is passed from the inlet b of the oily solution inlet channel and the inlet of the cell inoculation channel using a constant pressure pump c pass into the microfluidic chip and keep the air in the oil solution inlet channel 12 and the cell inoculation channel 13. The polyvinyl alcohol aqueous solution enters the aqueous solution inlet channel 11, the double-emulsion droplet buffer storage area 14, the double-emulsion droplet anchor area 15, the anchor droplet discharge channel 19, and the excess droplet discharge channels 16, 17 and 18, soaking The time is 15 minutes.

[0084...

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Abstract

The present invention relates to a micro-fluidic chip for observing and treating suspended cells in real time. The micro-fluidic chip comprises an upper substrate of a micro-fluidic channel and a lower substrate sealed with the upper substrate; the micro-fluidic channel in the upper substrate is composed of a double emulsion droplet generating area, a double emulsion droplet buffer storage area, adouble emulsion droplet anchoring area, an excess droplet discharging channel and an anchoring droplet discharging channel; and the lower substrate is composed of a glass piece and a polymer-materialfilm coating the surface of the glass piece. The micro-fluidic chip of the invention has good biocompatibility, thermal stability and gas permeability, is suitable for treating cells, and is suitablefor using a microscopic technology and a fluorescent staining technology to dynamically observe the cells in real time. The micro-fluidic chip of the present invention can precisely control the fluidby the driving force provided by anexternal pump, and realize the control of time and space for inputting suspension cells, a culture medium, a buffer solution and biochemical reagents used for treating the suspension cells. The micro-fluidic chip of the invention can realize single cell treatment and real-time observation on the suspension cells.

Description

technical field [0001] The present invention relates to the field of microfluidic chips, and specifically refers to a microfluidic chip for real-time observation and processing of suspended cells and its preparation method and application. The chip can coat suspended cells in double-emulsion droplets in the form of single cells Centered and fixed inside the chip to determine the position, it can perform real-time observation and biochemical treatment of a single suspension cell in batches. Background technique [0002] Microfluidic chip refers to a micro-unit fabricated and processed on a small area of ​​a substrate material through micro-nano processing technology, which can realize the biochemical treatment of samples. Microfluidic chip technology is characterized by a micron-scale fluid channel network, and uses special micro-nano structures to precisely control micro-liquids. On the one hand, it can integrate conventional biochemical laboratory detection and analysis t...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12M1/36
CPCC12M23/16C12M41/00
Inventor 王征王琳蔡博
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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