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Rare cell detection chip and application thereof

A rare cell and detection chip technology, applied in animal cells, tumor/cancer cells, vertebrate cells, etc., can solve the problems of poor selectivity and sensitivity, achieve the effect of enhancing selectivity or other characteristics and reducing additional pressure

Active Publication Date: 2014-07-23
GUANGZHOU ANFANG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invention solves the problem of poor selectivity and sensitivity in the rare cell capture process, can realize efficient and reliable rare cell capture and detection in a short time, and at the same time, the manufacturing process of the chip is simple, the degree of integration is high, the structure is dexterous and easy to operate. Simple, easy to apply, reliable and efficient

Method used

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  • Rare cell detection chip and application thereof
  • Rare cell detection chip and application thereof
  • Rare cell detection chip and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Such as Figure 1-6 Shown is a basic embodiment of a rare cell detection chip of the present invention. It includes a substrate 1 and a substrate 2 that is planarly consistent with the substrate 1 and sealed and bonded. The substrate 1 is composed of an upper layer 3, a lower layer 4 and a cell screening membrane 5 integrated between the upper layer 3 and the lower layer 4. One end of the upper layer 3 is provided with an upper layer sample inlet 6, and the other end is provided with an upper layer sample outlet 8. The upper layer sample inlet 6 is connected to an upper layer annular channel 12 with an upper layer gap 12.1 through the upper layer sample input channel 7, and the upper layer annular channel 12 passes through Several upper layer diffusion channels 14 that are radially arranged are connected to an upper layer circular sample pool 16, and this upper layer circular sample pool 16 is connected to the upper layer sample outlet 8 through the upper layer sample o...

Embodiment 2

[0066] Such as Figure 7 , 8 Shown is an embodiment of a rare cell detection chip of the present invention.

[0067] Figure 7 It is a scanning electron microscope image of the tapered micropore array filter membrane, wherein (a) is the side with the small diameter of the tapered micropores, and (b) is the side with the large diameter of the tapered micropores. The cell screening membrane 5 has a thickness of 40 micrometers. The thickness of the fixing ring 21 is 60 microns. The densely distributed and regularly arranged conical micropores on the cell screening membrane 5 have a small diameter of 6.5 microns, a large diameter of 25 microns, and a period of 40 microns. The material for making the microporous array filter membrane is polyethylene glycol.

[0068] Figure 8 for figure 1 The physical picture of the rare cell detection chip. The upper sample input channel 7, the upper sample output channel 8.1, the lower sample input channel 10, and the lower sample output ...

Embodiment 3

[0070] Using the rare cell detection chip in Example 2 to capture the colorectal cancer cell HT29 mixed in PBS solution includes the following steps:

[0071] 1) Sterilization: put the rare cell detection chip under ultraviolet light for sterilization;

[0072] 2) Surface modification of cell screening membrane 5: first inject 20 μl of MaxGel solution (diluted in PBS solution at a ratio of 1:100) into the chip with a syringe pump, incubate in the cell incubator for one hour, remove the residual solution, and then inject Phosphate buffered saline for washing;

[0073] 3) Cell capture: inject 10ml of phosphate buffer solution containing rectal cancer cell HT29 into the chip, so that the cancer cells are captured by the cell screening membrane 5;

[0074] 4) Cell fixation: inject a 4% paraformaldehyde solution into the chip to fix the captured cells on the cell screening membrane 5, and then inject phosphate buffer for washing;

[0075] 5) Cell staining: sequentially inject 0.2...

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Abstract

The invention discloses a rare cell detection chip and an application thereof. The chip comprises a matrix and a substrate, wherein the matrix is divided into an upper layer and a lower layer; a rare cell screening membrane is integrated between the upper layer and the lower layer; a sample inlet, a sample outlet and a sample channel are formed in each of the upper layer and the lower layer, which are suitable for the control of micro fluid such as blood samples, buffer solutions, cell culture solutions, other biochemical reagents and the like. When a sample is input from the sample inlet in the upper layer, and is output from the sample outlet in the lower layer, rare cells in the sample can be selectively blocked at the upper end of the cell screening membrane. According to the need, the buffer solutions, the cell culture solutions or other biochemical reagents are input from the sample inlet in the upper layer or the sample inlet in the lower layer, and are output from the sample outlet in the upper layer or the sample outlet in the lower layer so as to realize various cell operations and analysis and corresponding molecular biology detection. By adopting the rare cell detection chip disclosed by the invention, the problem of poor selectivity and sensitivity in capture of rare cells is solved; and the rare cell detection chip is suitable for efficient capture, culture and analysis of circulating tumor cells in the peripheral blood of the human body and fetal circulating cells.

Description

technical field [0001] The invention relates to a rare cell detection chip, which is suitable for the capture of circulating tumor cells and fetal circulating cells in human peripheral blood and related in vitro early diagnosis, auxiliary diagnosis, individualized treatment, chemotherapy drug evaluation, tumor recurrence monitoring and drug development etc. Background technique [0002] Cancer is the number one enemy of human health and the most common cause of death. Although there have been a lot of research and practice on the diagnosis and treatment of cancer, and many effective technologies and methods have been developed, these technologies and methods still have a lot of shortcomings in the two aspects of diagnosis and treatment. For example, we still can't do a good early diagnosis and evaluation of chemotherapy. Currently commonly used imaging techniques, including ultrasound (US), computed tomography (CT), magnetic resonance imaging (MRI) and positron emission co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12M1/12G01N15/10G01N1/30C12N5/09C12N5/00
CPCG01N15/00C12M45/00G01N15/01
Inventor 唐亚东石剑李思思汪莉陈勇
Owner GUANGZHOU ANFANG BIOTECH CO LTD
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