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Micro-fluidic chip integrating circulating tumor cell separation and single cell immunoblotting

A microfluidic chip and tumor cell technology, which is applied in the field of circulating tumor cell separation and protein analysis chips, can solve the problems of changes in cell function characterization, complex chip preparation process, poor biocompatibility, etc. The effect of fast speed and high separation rate

Active Publication Date: 2020-10-02
水熊健康科技(南通)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the preparation process of some chips is complicated, and it is difficult to control the separation and detection of single cells
The biocompatibility of the materials used in the preparation of the chip is not good enough, which may cause changes in the characterization of cell functions, so that the measured results cannot reflect the real situation in the real human life activity state

Method used

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  • Micro-fluidic chip integrating circulating tumor cell separation and single cell immunoblotting
  • Micro-fluidic chip integrating circulating tumor cell separation and single cell immunoblotting
  • Micro-fluidic chip integrating circulating tumor cell separation and single cell immunoblotting

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Example 1: Microchip Preparation.

[0104] 1. Take out a paper cup, put it on the balance, clear the balance, slowly pour 25g of polydimethylsiloxane (PDMS)) into the paper cup, clear the balance again, and slowly add 2.5g of polydimethylsiloxane (PDMS) with a 1mL pipette gun Polydimethylsiloxane curing agent The ratio of polydimethylsiloxane stock solution to polydimethylsiloxane curing agent is 10:1.

[0105] 2. Fully stir and mix the polydimethylsiloxane (PDMS) and polydimethylsiloxane (PDMS) curing agent with a stirring glass rod, and put them in a vacuum drying dish connected to a two-stage rotary vane vacuum pump. Apply vacuum for 10 minutes to ensure that there are no air bubbles in the mixed colloid. Pay attention to deflate in time during the vacuuming process to prevent the mixture from overflowing.

[0106] 3. Introduce the mixed colloid that has been evacuated into the petri dish where the silicon wafer with the target pattern is placed, so that the mixed ...

Embodiment 2

[0118] The breast cancer cell line MCF-7 cells are used to simulate circulating tumor cells in breast cancer patients, and MCF-7 cells are mixed into normal human blood to simulate blood samples collected from breast cancer patients to verify the various functional modules of the microchip .

[0119] Such as Figure 7 and 8As shown, when MCF-7 cells pass through the sorting module of the microchip, the video screenshot of cell distribution in the separation channel. The separation efficiency of MCF-7 was 68%, and the purity was 96%. Such as Figure 9 As shown, the purity of MCF-7 cells is higher after concentration and purification by membrane filtration. Such as Figure 10 As shown, the circulating tumor cells were captured by the cell trap, electrophoresed after in situ lysis, and then fixed by ultraviolet light irradiation. Protein analysis at the cellular level.

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Abstract

The invention discloses a microfluidic chip integrating circulating tumor cell separation and single cell immunoblotting. The microfluidic chip is characterized by comprising a circulating tumor cellsorting unit, a single cell capture unit, a photoactive gel electrophoresis separation unit and an immunoblotting analysis unit, wherein the circulating tumor cell sorting unit is used for separatingand sorting circulating tumor cells; the single cell capture unit is used for capturing single cells of sorted and enriched cells and performing closed lysis on the cells; the photoactive gel electrophoresis separation unit is used for pushing a protein obtained after cell lysis to a gel coating area on a chip; and the immunoblotting analysis unit is used for performing incubation and elution through a specific antibody, performing fluorescent or luminescent group labeling on a target protein molecule and detecting a signal of the target protein molecule. According to the invention, a varietyof functional units of cell sorting, purification, single cell capture, gel electrophoresis, immunoblotting analysis and the like of the circulating tumor cells are integrated on the micro-fluidic chip, so the separation speed, the analysis speed and the automation degree of the circulating tumor cells are greatly improved.

Description

technical field [0001] The invention relates to the field of circulating tumor cell separation and protein analysis chips, in particular to a microfluidic chip integrating label-free high-throughput circulating tumor cell separation and single circulating tumor cell immunoblotting. Background technique [0002] Circulating Tumor Cells (CTCs) [0003] With the improvement of people's quality of life and the extension of life expectancy, the mortality rate related to cancer has increased steadily in the past few decades, and cancer has become the number one killer of human beings. A study conducted by the World Health Organization found that at least 30% of cancer mortality could be prevented if cancer patients were diagnosed and treated before metastatic cancer occurred, which would lead to distant metastasis of the tumor along the blood circulatory system and death of the patient One of the main culprits is the circulating tumor cells (CTC) in the blood (1) . CTC refers t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12M1/42C12M1/12G01N33/68B01L3/00
CPCC12M23/16C12M29/04C12M35/02G01N33/68B01L3/5027B01L3/502707B01L3/502753B01L2200/10B01L2300/12
Inventor 丁显廷阿依努尔·阿卜拉张婷谢海洋
Owner 水熊健康科技(南通)有限公司
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