Micro fluidic device for CTC high-flux unicell phenotype analysis

A single-cell, microfluidic chip technology, used in measurement devices, material analysis by optical means, material analysis, etc., can solve the problems of rapid and continuous sampling of single cells, slow application development, etc. effect of change

Active Publication Date: 2019-06-28
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in terms of rapid and continuous single-cell analysis of CTCs, the application development is slow due to the trouble of rapid and continuous single-cell injection of rare CTCs.

Method used

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  • Micro fluidic device for CTC high-flux unicell phenotype analysis
  • Micro fluidic device for CTC high-flux unicell phenotype analysis
  • Micro fluidic device for CTC high-flux unicell phenotype analysis

Examples

Experimental program
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Effect test

Embodiment 1

[0067] Such as figure 1 As shown, the integrated microfluidic device of the present disclosure was used to perform single-cell phenotype analysis on colorectal cancer patients with different clinical stages. First, take 5 mL of fresh anticoagulated blood, and after being treated with erythrocyte lysate, the red supernatant is discarded by centrifugation, and the white precipitate at the bottom is redispersed in 5 mL of PBS. Liquid PBS is passed into the second inlet at 750uL / min. At the outlet of the spiral chip unit, the inner third outlet is connected to the fourth inlet of the microcolumn chip unit through a catheter. The outer outlet is the waste liquid outlet. min leads to the fifth inlet of the micro-column chip unit, and the tumor cells can further concentrate the liquid through the lateral deviation of the micro-column to improve the purity of the tumor cells. The sixth outlet of the microcolumn chip is the waste liquid outlet, and the seventh outlet is the collection...

Embodiment 2

[0069] The established integrated chip device was used to count and phenotype CTCs in blood samples. As controls, blood collected from nine healthy donors was also analyzed in the same way. Judging from the test results ( Figure 7 a), The number of CTCs in patient blood samples is closely related to tumor stage. The average number of CTCs in patients with clinical stage 2 to 4 tumors was 55, 125 and 199, respectively, while the number of CTCs detected in the control group was less than 20. In addition, the present disclosure found that with the aggravation of the degree of tumor, the expression of EpCAM in CTCs decreased and the expression of N-cad increased ( Figure 7 b, c), illustrating that more CTCs are in the mesenchymal phenotype with higher metastatic ability and invasive ability, which means stronger tumor metastatic ability. The expression of two proteins in CTCs of patients with different stages Figure 7 There are obvious differences in location distribution i...

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Abstract

The invention relates to a micro fluidic device for CTCs selecting and gathering and unicell analysis, belongs to the technical field of micro fluidic chip detection, and aims at solving the technicalproblem that CTC phenotype analysis is difficult in clinic application due to factors including low cell concentration in CTCs, large cell phenotype difference, low unicell analysis flux and nonsystematic automation in a sample. According to the micro fluidic chip device for rapid CTCs selection and high-flux unicell analysis, a spiral chip and micro-column array lateral flow chip are used to complete selecting and gathering of tumor cells, and the micro-column array chip carries out phenotype analysis detection after EpCAM and N-cad marking,. All operations for CTCs are implemented in a micro-nano scale platform CTCs loss is reduced to largest degree, the method has the advantages of high integrated degree, high flux, convenient operation integration and automation, and the method can beapplied to liquid biopsy in clinics conveniently, rapidly and easily.

Description

technical field [0001] The disclosure relates to the technical field of microfluidic chip detection, in particular to a system-integrated microfluidic chip device for rapid sorting of CTCs and continuous on-line single-cell analysis, especially for loss-free acquisition and monitoring of all CTCs in blood samples. Sequential high-throughput single-cell analysis. Background technique [0002] The information disclosed in this Background section is only intended to increase the understanding of the general background of the disclosure, and is not necessarily to be taken as an acknowledgment or any form of suggestion that the information constitutes prior art that is already known to those skilled in the art. [0003] Tumor metastasis relies on the release of circulating tumor cells (CTCs) from the primary tumor, which migrate to distant tissues and form secondary tumors. CTCs carry a large amount of molecular information of the primary tumor and present dynamic changes in the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/01G01N21/64B01L3/00
Inventor 唐波李璐裴海盟司海滨
Owner SHANDONG NORMAL UNIV
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