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Circulation tumor cell automatic capturing micro-fluidic chip and automatic capturing method thereof

A microfluidic chip, automatic capture technology, applied in the fields of molecular biology and medicine, can solve the problems of no biological activity, high false positive rate, and inability to detect CTCs, so as to reduce human intervention, prevent clogging, and continuously and effectively sorting effect

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

Although Cellsearch has been approved by the FDA, there are still some defects in the system, such as the inability to realize automatic sorting, high false positive rate, enriched CTCs have no biological activity, etc. More importantly, the CTCs captured by Cellsearch are only EpCAM-positive types, and many highly malignant CTCs do not express EpCAM, so they cannot be detected

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] See eg 1 and figure 2 As shown, this embodiment provides a microfluidic chip for automatic capture of circulating tumor cells. A microfluidic chip for automatically capturing circulating tumor cells includes a central control module 10 , a sample injection module 20 , a sample sorting module 40 and a buffer solution injection module 30 .

[0045] see figure 2 As shown, the sample sampling module 20 includes a sampling unit 21 and a sampling microchannel 22 . The buffer sampling module 30 includes an oscillatory flow unit 31 and a continuous flow unit 32 . The sample sorting module 40 includes a sample sorting unit 44 , a target cell collection unit and a non-target cell collection unit 46 .

[0046] The sampling unit 21 , the sampling microchannel 22 and the sample sorting module 40 are connected in sequence, and the sampling microchannel 22 communicates with the accommodating cavity 441 of the sample sorting unit 44 . The central control module 10 is connected to...

Embodiment 2

[0061] In this example, the microfluidic chip for automatic capture of circulating tumor cells in Example 1 is used to detect clinical samples.

[0062] The microfluidic chip for automatic capture of circulating tumor cells in Example 1 was used to detect peripheral blood samples (numbered 1-20) of 20 tumor patients.

[0063] 1. Reagent formula:

[0064] (1) Preparation of blocking solution: minimum essential medium (MEM) contains 15% polysucrose, 0.2% Pluronic and 5% bovine serum albumin (BSA).

[0065] (2) Hank's balanced salt solution:

[0066]

[0067] 2. Specific detection steps:

[0068] (1) Sealing: the sample sorting unit 44 of the sample sorting module 40 is made of the bottom sheet of PDMS and the upper cover glass sheet, in order to prevent non-specific adsorption of cells and PDMS and glass surfaces, before using the sample sorting module 40, It needs to be closed. Inject the blocking solution prepared above through the buffer inlet and the sample inlet, so ...

Embodiment 3

[0077] In this example, the microfluidic chip for automatic capture of circulating tumor cells in Example 1 was used to detect cell lines.

[0078] 1. Sample preparation

[0079] 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 normal human peripheral blood, and prepare samples with cell concentration gradients as shown in the following table:

[0080] Table 2 Sample Cell Concentration

[0081]

[0082] 2. Sample testing

[0083] Using the circulating tumor cell automatic capture microfluidic chip in Example 1 and the detection method in Example 2, sample numbers 21-30 were detected, and the enriched e...

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Abstract

The invention discloses a circulation tumor cell automatic capturing micro-fluidic chip and an automatic capturing method thereof. The circulation tumor cell automatic capturing micro-fluidic chip comprises a sample feeding module, a sample separation module and a buffer liquid sample feeding module, wherein the sample feeding unit is connected with the sample separation module; the sample feeding unit is used for feeding a sample to be detected into the micro-fluidic chip; the buffer liquid sample feeding module comprises an oscillation flow unit and a continuous flow unit; the oscillation flow unit and the continuous flow unit are respectively connected with the micro-fluidic chip; the sample separation unit is used for separating cells of the sample to be detected; a target cell collection unit is used for collecting target cells obtained through separation; the micro-fluidic chip comprises a sample separation unit, a target cell collection unit and a non-target cell collection unit; and the non-target cell collection unit is used for collecting non-target cells and generated waste liquids obtained after separation. The circulation tumor cell automatic capturing micro-fluidic chip is capable of achieving automatic separation of cells and is low in false positive rate.

Description

technical field [0001] The invention relates to the fields of molecular biology and medicine, in particular to a microfluidic chip for automatically capturing circulating tumor cells and an automatic capturing 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. However, the amount of circulating tumor cells in peripheral blood is very small, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12N5/09
CPCC12M23/16C12M33/08C12M33/12C12M47/04C12N5/0693
Inventor 刘志明吴诗扬廖传荣
Owner SUREXAM BIO TECH
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