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Micro-fluidic chip used for cell sorting and gathering and application of micro-fluidic chip

A microfluidic chip and sorting technology, applied in the field of modern medicine, can solve the problems of inability to study cell biological characteristics, restrict cell purity, lack of cell collection or enrichment, etc., and achieve good cell capture efficiency, good cell activity, and convenient The effect of scientific research and disease evaluation analysis

Active Publication Date: 2016-04-13
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The above strategies have some limitations in the sorting and enrichment of CTCs. For example, when using method (1) for cell sorting, the specificity is not high, which seriously restricts the purity of the cells captured by this method; due to the friction of magnetic beads and cell invasion However, the cells obtained by method (2) are often dead cells, which can only be used to count the number of cells, but cannot further study the biological characteristics of cells; method (3) is used for relatively large numbers in enriched environments It is an effective means for target cells, but for relatively small number of cells such as CTCs, even if they can be separated from non-target cells, there is no means to collect or enrich the cells

Method used

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  • Micro-fluidic chip used for cell sorting and gathering and application of micro-fluidic chip
  • Micro-fluidic chip used for cell sorting and gathering and application of micro-fluidic chip
  • Micro-fluidic chip used for cell sorting and gathering and application of micro-fluidic chip

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Experimental program
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Effect test

Embodiment 1

[0090] The preparation of embodiment 1 chip body

[0091] The preparation process and working principle of the chip body are as follows: figure 1 Shown, its preparation process comprises the following steps:

[0092] (1) Use dichloromethane, acetone, methanol and deionized water to ultrasonically clean the ITO conductive glass for 15 minutes, then use plasma etching (Harrick PCD32G) to treat the ITO conductive glass for 5 minutes, and put the treated ITO conductive glass into the pre-prepared Stand still 3min in the chitosan solution;

[0093] (2) According to the standard method of layer-by-layer self-assembly, soak the ITO conductive glass alternately in the chitosan solution and the Prussian blue nanoparticle solution for layer-by-layer self-assembly treatment. Wash with .0 deionized water for 3 minutes to form a micro-voltage response release layer (CHI / PB) on the surface of ITO conductive glass 27 ;

[0094] (3) According to the standard method of layer-by-layer self-...

Embodiment 2

[0098] Example 2 Preparation of Cell Sorting and Enrichment Device

[0099] Such as Figure 2a As shown, a cell sorting and enrichment device in this embodiment includes a microfluidic chip 1, a micro-injection pump 2, a three-way tube 3, a waste liquid collection tube 4, a target cell collection tube 5, and an electrochemical workstation (Fig. omitted).

[0100] Depend on Figure 2a and 2b It can be seen that the microfluidic chip 1 includes a stacked and fixed chip body 12 (made by Example 1) and a cover 11. The cover 11 is provided with a groove 13, a sample injection channel 14 communicated with the groove 13, and a The sample outlet channel 15 , and the opening of the groove 13 faces the chip body 12 .

[0101] The cover body 11 is made of PDMS material, and it is convenient to set the groove 13 , the sample inlet channel 14 and the sample outlet channel 15 thereon. The fixing method of the cover 11 and the chip body 12 is as follows: when the cover 11 is relatively ...

Embodiment 3

[0108] Embodiment 3 morphology characterization and performance test

[0109] (1) Carry out morphology characterization to the product obtained in embodiment 1 step (2)

[0110] Figure 3a Shown is the appearance diagram of the corresponding product when n in the micro-voltage response release layer (CHI / PB)n is 0, 3, 6, 9, 12, 15, 18, 21, 24, and 27 respectively.

[0111] Use ellipsometry Puyi to measure the film thickness of products with different number of layers, the results are as follows Figure 3b shown.

[0112] Utilize the ultraviolet-visible spectrophotometer to detect the linear change of the absorption value at 700nm of the products with different treatment layers, the results are as follows Figure 3c shown.

[0113] Utilize the scanning electron microscope to obtain the product surface topography map of n=27, such as Figure 3d shown.

[0114] Utilize the atomic force microscope to obtain the product surface topography map of n=27, such as Figure 3e show...

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Abstract

The invention discloses a micro-fluidic chip used for cell sorting and gathering and an application of the micro-fluidic chip. The micro-fluidic chip comprises a chip body and a cover, which are fixed in overlaying arrangement. The cover is provided with a groove, and a sample feeding channel and a sample discharging channel, which communicate with the groove. An opening of the groove faces the chip body. The chip body is obtained through the following method comprising: (a) performing self-assembly of chitosan / prussian blue nanoparticles layer by layer on the surface of a conductive substrate; (b) performing self-assembly of polydiallyldimethylammonium chloride / polyacrylic acid layer by layer on the surface of the product obtained from the step (a); and (3) performing carboxyl-group activating treatment, avidin modification, and biotinylation antibody coupling of the product obtained from the step (b). An environmental-stimulus responding material is introduced to the micro-fluidic chip, so that properties of the micro-fluidic chip are changed in the function of outside stimulus, and the micro-fluidic chip can control and losslessly release captured cells.

Description

technical field [0001] The invention belongs to the technical field of modern medicine, and in particular relates to a microfluidic chip for cell sorting and enrichment and its application. Background technique [0002] Cancer is the number one cause of death in developed countries and the second in developing countries. Clinically speaking, the main reason for the high mortality of cancer patients is the delay of disease diagnosis and inappropriate treatment plan. Physiologically, most of the deaths caused by cancer are caused by the migration of tumor cells to other organs in the body through the hematogenous route. In this process, metastatic cancer cells, namely circulating tumor cells (CTCs), play a key role in cancer metastasis. Accumulating evidence indicates that the metastasis of tumor cells from the primary site to other sites occurs early in cancer development. Therefore, the collection and biochemical analysis of CTCs can provide an effective channel for the e...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N35/00
Inventor 王本肖云唐睿康
Owner ZHEJIANG UNIV
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