Microfluidic chip integrating circulating tumor cell capture, lysis and nucleic acid detection, and device and method of microfluidic chip

A microfluidic chip and microfluidic device technology, applied in the field of microfluidics, can solve the problems of high cost, cumbersome process, and the inability to simultaneously capture, lyse, and nucleic acid detection of circulating tumor cells, so as to reduce flow resistance and integrate High degree of automation, improved detection speed and sensitivity

Active Publication Date: 2019-06-07
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a microfluidic chip and its device and method that integrate the capture, lysis and nucleic acid detection of circulating tumor cells, so as to solve

Method used

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  • Microfluidic chip integrating circulating tumor cell capture, lysis and nucleic acid detection, and device and method of microfluidic chip
  • Microfluidic chip integrating circulating tumor cell capture, lysis and nucleic acid detection, and device and method of microfluidic chip
  • Microfluidic chip integrating circulating tumor cell capture, lysis and nucleic acid detection, and device and method of microfluidic chip

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Example 1 Preparation of microfluidic chip

[0041] 1.1 First, for the structural part of the microfluidic chip 100, use AutoCAD drawing software to draw the required structural graphics, make a mask plate, and use a four-inch single crystal silicon wafer as the substrate for thermal oxidation to form a 2 μm oxide layer. Then perform glue-coating photolithography, reactive ion etching, glue-removing cleaning, glue-coating photolithography, and deep reactive ion etching to etch out the height of the filter channel in the circulating tumor capture area. Then remove the photoresist and perform deep reactive ion etching according to the silicon dioxide pattern to etch the height of other channels of the chip and the depth of the nucleic acid microcavity, and finally obtain a silicon wafer mold with a microstructure .

[0042] 1.2 Place the silicon wafer mold and the open centrifuge tube containing 10 μL of fluorosilane in a vacuum drying oven, vacuum to negative pressure t...

Embodiment 2

[0052] Embodiment 2 Assembly of microfluidic device

[0053] Such as Figure 5 Shown is a microfluidic device provided according to a preferred embodiment of the present invention, the microfluidic device includes: a solution storage chamber 200 connected in sequence, a microfluidic chip 100, a waste liquid collection syringe 300, and a sample injection The pump 400 also includes an air pump 500 and a nitrogen tank 600 connected to the T-shaped pipe structure 9 of the valve control area 30 of the microfluidic chip 100 . Wherein, the solution delivery needle tube 700 is passed between the microfluidic chip 100 , the solution storage chamber 200 and the waste liquid collection syringe 300 .

[0054] It should be understood that the intake mechanism is not limited to the air pump 500 and the nitrogen tank 600 used in this embodiment, but can also be any device that can be connected to the T-shaped pipeline structure 9 and supply gas to the second pipeline 92 .

Embodiment 3

[0055] Example 3 Capture, Lysis and Nucleic Acid Amplification Detection of Circulating Tumor Cells Using a Microfluidic Device

[0056] Firstly, the blood samples were processed by using Ficoll-Paque plus reagent, and then the cells were fixed with 4% paraformaldehyde, centrifuged to remove the supernatant, resuspended with 1mL PBS, and placed in a 4°C refrigerator for later use. During detection, the air pressure pump 500 is first turned on to provide air pressure to close the sampling channel 6, then the flow rate of the sampling pump 300 is set to 10mL / h, the diameter is set to 19mm, and one end of the waste liquid collection syringe 300 is connected to the microfluidic chip 100 The cell capture sample outlet 5 of the microfluidic chip 100 is connected to the negative pressure provided by the sample pump 300 to draw the sample into the microfluidic chip 100 from the cell capture inlet 1 of the microfluidic chip 100, and the smaller white blood cells and red blood cells Tha...

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Abstract

The invention provides a microfluidic chip integrating circulating tumor cell capture, lysis and nucleic acid detection, and a device and method of microfluidic chip. The microfluidic chip is formed by laminating a glass base layer and a polydimethylsiloxane chip layer. The polydimethylsiloxane chip layer comprises the following parts: a circulating tumor cell capture zone; a nucleic acid detection zone comprising a feeding channel with a flexible sidewall, a microcavity array region and a sample outlet; and a valve control region comprising: T-shaped pipe structures respectively disposed on both sides of the feeding channel, wherein the structure includes a first pipe extending parallel to the feeding channel and a second pipe vertically connected to the feeding channel, and the second pipe is connected to a gas supply mechanism to control the opening and closing of the feeding channel by controlling the opening and closing of the gas supply mechanism. According to the present invention, the microfluidic chip integrating the circulating tumor cell capture, lysis and nucleic acid detection, and the device and method have the advantages of high throughput, low cost, high sensitivity, easy usage, and short time consumption.

Description

technical field [0001] The invention relates to the field of microfluidics, and more specifically relates to a microfluidic chip, a device and a method thereof that integrate capture, lysis and nucleic acid detection of circulating tumor cells. Background technique [0002] Worldwide, the incidence of cancer has remained high for a long time, and it is the main cause of death in cancer patients. The same is true in China, regardless of rural cities, regardless of men and women, cancer is still one of the diseases with a very high mortality rate, and has become a major issue affecting public health. The key to fighting cancer is early diagnosis and treatment of primary cancer. [0003] Circulating tumor cells (CTCs) are tumor cells that have shed from the primary tumor lesion and invaded the peripheral blood circulation. Since CTCs are closely related to the development and metastasis of cancer, and besides containing genetic material similar to the primary tissue, CTCs also...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12M1/36C12M1/34C12M1/04C12Q1/6851
Inventor 贾春平王艳敏郜晚蕾金鎏赵辉齐同周洪波赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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