Circulation tumor cell automatic separation and purification micro-fluidic chip and separation and purification method thereof

A microfluidic chip and automatic separation technology, applied in the field of biomedicine, can solve problems affecting CTCs protein and molecular research, achieve the effect of accurate critical separation size, avoid cell damage, and avoid false results

Active Publication Date: 2017-11-28
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some blood cells in the blood are large in size and overlap with the size of CTCs. Therefore, the CTCs enriched by DLD technology still contain a large number of large blood cells, which affects the subsequent research on CTCs proteins and molecules.

Method used

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  • Circulation tumor cell automatic separation and purification micro-fluidic chip and separation and purification method thereof
  • Circulation tumor cell automatic separation and purification micro-fluidic chip and separation and purification method thereof
  • Circulation tumor cell automatic separation and purification micro-fluidic chip and separation and purification method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] This embodiment provides a microfluidic chip 10 for automatic separation and purification of circulating tumor cells.

[0041] see figure 1 As shown, a microfluidic chip 10 for automatic separation and purification of circulating tumor cells includes a separation and purification chamber 100 , a separation column 200 , a microcolumn 300 and a target cell collection chamber 600 . The microfluidic chip 10 for automatic separation and purification of circulating tumor cells is denoted as microfluidic chip A for automatic separation and purification of circulating tumor cells.

[0042] Automatic Separation and Purification of Circulating Tumor Cells The separation and purification chamber 100 of the microfluidic chip 10 is formed by bonding a bottom sheet and an upper cover. The material of the bottom sheet includes but not limited to: glass, PDMS, silicon material, etc.; the material of the upper cover includes but not limited to: glass, PDMS, silicon material, etc. The ...

Embodiment 2

[0059] Using the microfluidic chip A for the automatic separation and purification of circulating tumor cells in Example 1 to detect clinical samples

[0060] In this example, the microfluidic chip A for automatic separation and purification of circulating tumor cells in Example 1 was used to detect peripheral blood samples (numbered 1-20) of 20 tumor patients.

[0061] 1. Reagent formula:

[0062] (1) Preparation of blocking solution: minimum essential medium (MEM) contains 15% polysucrose, 0.2% Pluronic and 5% bovine serum albumin (BSA).

[0063] (2) Hank's balanced salt solution:

[0064]

[0065] 2. Specific detection steps:

[0066] (1) Closed: The separation and purification chamber of the microfluidic chip 10 for automatic separation and purification of circulating tumor cells is composed of a bottom sheet of PDMS and an upper cover glass sheet, in order to prevent non-specific contact between cells and PDMS and the glass surface of the separation and purification ...

Embodiment 3

[0075] Using the microfluidic chip A for the automatic separation and purification of circulating tumor cells in Example 1 to detect cell lines

[0076] 1. Sample preparation

[0077] In order to prove that the microfluidic chip A for automatic separation and purification of circulating tumor cells of the present invention has a good capture rate, can effectively detect tumor cells in the sample, and remove normal blood cells in the sample at the same time, the number of tumor cells in this example has been Known samples were taken as examples for testing. In this embodiment, the epithelial cell line MCF-10A, the mesenchymal tumor cell line U118, the epithelial-mesenchymal mixed lung cancer cell line PC-9 and the negative control CCRF-HSB-2 lymphoblastoid cells were selected for the experiment. Those skilled in the art As long as you know the name of the cell line, you can buy it. Use a cell counter to take a certain number of the above-mentioned cells, add them to 5 mL of n...

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Abstract

The invention discloses a circulation tumor cell automatic separation and purification micro-fluidic chip and a separation and purification method thereof. The circulation tumor cell automatic separation and purification micro-fluidic chip comprises a separation and purification chamber, a diversion column and micro columns, wherein a sample inlet, a first outlet from which target cells flow out, and a plurality of second outlets from which non-target cells flow out are formed in the separation and purification chamber; the separation and purification chamber is internally provided with two micro column arrays; each micro column array consists of a plurality of micro column rows; each micro column row comprises a plurality of micro columns; a gap is formed between every two adjacent micro columns of one micro column row; the gap forms a non-target cell channel; a gap is formed between two micro column arrays; the gap forms a target cell channel from which the target cells pass; the width of the target cell channel is greater than that of the non-target cell channel; an acute angle is formed between the micro column rows and the target cell channel; and the acute angle faces to the sample inlet. The circulation tumor cell automatic separation and purification micro-fluidic chip is high in separation and purification degree.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a microfluidic chip for automatic separation and purification of circulating tumor cells and a separation and purification method thereof. Background technique [0002] Circulating Tumor Cells (CTCs) in human peripheral blood are a type of rare cells that are rare in number but have important clinical significance. Under certain conditions, it can develop into tumor metastatic lesions. In general, circulating tumor cells indicate the presence of tumors in humans and possible metastasis. Since more than 90% of clinical cancer deaths are caused by metastasis, and circulating tumor cells provide a source of tumor metastatic lesions in the human body other than tumor lesions, therefore, capturing and detecting circulating tumor cells from blood is becoming more and more important. draw people's attention. [0003] However, the amount of circulating tumor cells in peripheral blood is ver...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/574
Inventor 刘志明吴诗扬廖传荣
Owner SUREXAM BIO TECH
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