A modular chamber and its application for efficient capture of rare cells in blood

A modular and cell-based technology, applied in the field of biofunctional materials, analysis and detection, can solve the problems of low separation purity, small processing capacity, and inability to simultaneously realize large-capacity and high-efficiency capture of rare cells to be tested by physical methods, and achieve the preparation method Simple and fast, fast capture, overcome the effect of small amount of blood processing

Active Publication Date: 2020-12-11
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Based on the technical problems that the processing capacity of the microfluidic method is too small, the separation purity of the physical method is low, and the rare cells to be tested cannot be captured at the same time with large capacity and high efficiency, the present invention proposes a new design idea of ​​modular chambers

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  • A modular chamber and its application for efficient capture of rare cells in blood
  • A modular chamber and its application for efficient capture of rare cells in blood
  • A modular chamber and its application for efficient capture of rare cells in blood

Examples

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preparation example Construction

[0033] The preparation method of the modular chamber for efficient capture of large blood volume circulating tumor cells of the present invention comprises the following steps:

[0034] (1) Prepare two plates of polymer material or inorganic material, respectively as the upper sealing plate 1 and the lower sealing plate 2; and respectively set the liquid outlet 7 and the liquid inlet 8 on their surfaces. The liquid is injected from the liquid inlet 8 of the lower sealing plate, and the small chamber is filled in a “snake shape” from bottom to top. After being statically captured for a period of time, it flows out from the liquid outlet 7 of the upper sealing plate for subsequent detection.

[0035] (2) Several CTCs capture substrates 4 are prepared. The substrates have high-efficiency CTCs capture performance, and their size should be slightly smaller than that of the upper sealing plate 1 and the lower sealing plate 2 .

[0036] (3) Prepare the same number of ring gaskets 6 a...

Embodiment 1

[0045] In this embodiment, plexiglass is used as the upper sealing plate 1 and the lower sealing plate 2 respectively, and the inlaid syringe needles are used as the liquid outlet 7 and the liquid inlet 8, and the CTCs capture substrate 4 is the surface of the polydimethylsilane (PDMS) for cell replication. Screws are assembled to prepare modular chambers; by examining the capture efficiency of modular chambers for different types of cancer cells, the capture system of the present invention is further described and verified, including the following steps:

[0046] 1) Preparation of the modular chamber:

[0047] First prepare two plexiglass planes of 60mm×40mm as the upper sealing plate 1 and the lower sealing plate 2 respectively, and insert the syringe needle into them as the liquid outlet 7 and the liquid inlet 8 . Then prepare several 50mm×30mm CTCs capture substrates 4 and the same number of 40mm×25mm×3mm “back” shaped rubber gaskets 6 . Each CTCs capture substrate 4 is p...

Embodiment 2

[0058] In this example, the surface of the cell is selected as the CTCs capture substrate of the modular chamber, and different numbers of MCF-7 cells are added to the whole blood as the CTCs artificial blood sample model to be captured; and the captured cancer cells and white blood cells are scanned Observation by electron microscope further elaborates and verifies the capture system of the present invention, comprising the following steps:

[0059] 1) The preparation method of the modular chamber is the same as in Example 1.

[0060] 2) The preparation method of the CTCs capture substrate 4 is the same as that in Example 1.

[0061] 3) Prepare circulating tumor cell samples to be tested:

[0062] Here, MCF-7 cells were pre-stained with DAPI dye to make them fluoresce blue. Take 100 μL of pre-stained MCF-7 cell suspension, count with a cell counter, and dilute to 1×10 with DMEM cell culture medium. 3 The concentration of each cell / mL is ready for use; take 9 mL of fresh an...

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Abstract

The invention discloses a modularized small chamber and application thereof to efficient capturing of rare cells in blood. The modularized small chamber is composed of an upper sealing plate, a lowersealing plate and at least one containing cavity module, wherein the bottom surface of the containing cavity module is a cell capturing substrate; a liquid through hole is formed in the upper sealingplate and / or the lower sealing plate. The containing cavity module is composed of the detachable cell capturing substrate and an annular gasket capable of being matched with the cell capturing substrate to use. The modularized small chamber can be combined with any one cell capturing substrate, realizes efficient and specific capturing of the rare cells with different types and has a wide application range. A method has a simple manufacturing technology and low cost, and a complicated surface etching technology is not needed. Furthermore, the treatment amount of the blood can be regulated andcontrolled through regulating and controlling the quantity of the modules; the modularized small chamber is flexible and convenient when being used for coping with rare cell blood samples with different capacities. The modularized small chamber is hopefully used for early diagnosis of diseases of patients, and is especially applicable to the efficient capturing of large-blood-volume CTCs samples of patients with tumors.

Description

technical field [0001] The invention belongs to the technical field of biological functional materials and analysis and detection, and in particular relates to the preparation of an analysis and detection device of a modular small chamber, which is used to capture rare cells in blood, especially the technology of circulating tumor cells. Background technique [0002] Some cells in the blood are present in very small amounts but are important for understanding disease, such as stem cells, circulating endothelial cells, residual disease cells, and circulating tumor cells. Accurate detection and analysis of these rare cells is critical to understanding disease processes and pathogenesis, especially circulating tumor cells are closely related to early diagnosis of cancer. [0003] Circulating tumor cells (CTCs) refer to tumor cells that have shed from solid tumor lesions, undergo epithelial-mesenchymal transition (EMT), and thus have flow characteristics and enter the peripheral...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/574G01N33/68C12M1/00
CPCC12M47/04G01N33/56966G01N33/574G01N33/6893G01N2800/7028
Inventor 贾凌云韩璐璐孙贺杨立为
Owner DALIAN UNIV OF TECH
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