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High-flux cell sorting and enriching device and using method thereof

A high-throughput, cell-based technology, applied in the biological field, can solve the problems of cell screening barriers in yield, difficult single cell tracking and observation, and low enrichment purity, etc., to achieve continuous culture and proliferation, low shear force, and integration Effect

Active Publication Date: 2020-08-07
INST OF MICROELECTRONICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been applied to the removal of large particle impurities and the separation of large and small cells in cell suspensions, and it also has a good effect on the enrichment of tumor cells. The disadvantages of low concentration and low enrichment purity due to easy clogging of pores during the filtration process limit its application in cell biology research and clinical diagnosis; in addition, various cost factors such as yield rate in practical applications are also important factors in the promotion of cell screening technology. an important obstacle
[0003] For the follow-up detection of recovered cells, traditional biological cell experiments are mostly completed in well plates, and then directly dropped on glass slides for observation, which is time-consuming and laborious. Reagents A large amount is used, and it is difficult to track and observe single cells and maintain a uniform microenvironment

Method used

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  • High-flux cell sorting and enriching device and using method thereof
  • High-flux cell sorting and enriching device and using method thereof
  • High-flux cell sorting and enriching device and using method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Please refer to Figure 4 with Figure 5 , for the high-throughput rare cell enrichment device based on the inclined filter membrane, the role of the channel structure 6 is not only to support the cell filter membrane 4 to provide channels, but also to cause disturbance to the fluid, so that the cells are in a state of flushing instead of Will adhere to the surface of the cell filter 4, such as Image 6 As shown, microfluidic channels 8 of different shapes can achieve this effect.

[0047] The operation method of the high-throughput rare cell enrichment device using the integrated single cell ( / cell cluster) culture observation module of this embodiment includes the following steps:

[0048](1) Experiment preparation. Firstly, pretreat the sample obtained by liquid biopsy, such as blood dilution, primary filtration of pleural effusion, etc., select a certain amount of biopsy cell sample according to actual needs, take 1-30ml of the volume, and complete the packaging a...

Embodiment 2

[0053] In this example, please refer to Figure 7 Similar to Example 1, it also includes a three-layer sandwich enrichment device consisting of a cell filter 4, a channel structure 6 above the membrane and a support structure 5 below the membrane, and the difference is that the cell filter 4 can be fixed in multiple layers; Figure 7 Shown is the immobilization of two layers of cell filters 4, the size of the upper filter membrane microwell array is larger, which can keep the large cells 4 above the upper filter membrane, and the medium and small cells 11 flow into the upper channel of the second layer filter membrane , to achieve the enrichment of medium cells again, and finally the small cells 11 flow out through the second layer of filter membrane, and the channel formed by the support structure 5 under the membrane and the second layer of cell filter membrane 4 realizes washing and cell recovery through PBS buffer 13, the principle is the same as The monolayer enrichment d...

Embodiment 3

[0060] In this example, please refer to Figure 8 Similar to Example 1, it also includes a three-layer sandwich enrichment device composed of a cell filter 4, a channel structure 6 and a support structure 5 below the membrane. The difference is that the cell filter 4 bends in the tortuous flow channel and is formed by the cell filter 4 itself The non-planar flow channel of the membrane causes the disturbance of the fluid in the channel above the membrane, which accelerates the process of cell enrichment.

[0061] The operation method of the high-throughput cell sorting and enriching device of this embodiment includes the following steps:

[0062] (1) Experiment preparation. Firstly, pretreat the sample obtained by liquid biopsy, such as blood dilution, primary filtration of pleural effusion, etc., select a certain amount of biopsy cell sample according to actual needs, take 1-30ml of the volume, and complete the packaging and processing of the device. After disinfection, firs...

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Abstract

Aiming at the technical defects of low target cell screening flux, large damage, easy loss and pollution of recovered cells and the like in the existing cell research technology, the invention provides a high-flux cell sorting and enriching device and a using method thereof. The device has the following advantages: 1) cell enrichment, recovery, capture and culture are integrated in a closed mode,so the problems of inter-link cell loss and pollution are solved; 2) on a cell separation module, by combining an inclined cell filter membrane and a fluid channel, the target cell flux is kept on thebasis of keeping the cell activity; and 3) a single cell ( / cell cluster) capture and culture observation module solves the problem of cell tracking and positioning detection, and effectively culturesproliferative cells. The device is simple and convenient to operate, and provides equipment technical support for preparation and amplification of rare cell samples and single cell accurate researchin biomedicine and clinical diagnosis research.

Description

technical field [0001] The invention belongs to the field of biotechnology, and particularly provides a high-throughput cell sorting and enriching device and its application method. Background technique [0002] Cells are the basic unit of living organisms. Tissues or body fluids of higher organisms (such as the human body) are composed of a variety of mixed cells. Separating and purifying target cells from mixed cell samples is an important step in current cell research. For biological research It is of great significance for both clinical and diagnostic analysis (such as tumor liquid biopsy). The detection and accurate evaluation of enriched tumor cells is clinically important, and can guide the staging and clinical management of cancer patients; currently, the clinical diagnosis of tumor cells is mainly through cytological analysis and invasive biopsy methods. The sensitivity of the method depends largely on the amount and quality of the fluid and the experience of the c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/12C12M1/34C12M1/00
CPCC12M33/14C12M47/04C12M23/16C12M23/42C12M41/36
Inventor 赵阳程洁黄成军
Owner INST OF MICROELECTRONICS CHINESE ACAD OF SCI
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