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Combined sorting purification device and sorting detection method for circulating blood tumor cells

A tumor cell and purification device technology, applied in the field of combined sorting and purification of circulating blood tumor cells and sorting and detection, can solve the problems of high cost, poor specificity, low sensitivity, etc. small device effect

Active Publication Date: 2017-05-31
北京方远智汇科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to overcome the problems of high cost, low sensitivity, and poor specificity of the prior art CTC sorting and purification technology, the present invention combines microfluidic chip technology and immunomagnetic bead sorting technology to provide a combined sorting of circulating blood tumor cells Purification device and method for detecting circulating blood tumor cells

Method used

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  • Combined sorting purification device and sorting detection method for circulating blood tumor cells
  • Combined sorting purification device and sorting detection method for circulating blood tumor cells
  • Combined sorting purification device and sorting detection method for circulating blood tumor cells

Examples

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Effect test

Embodiment 1

[0058] Example 1 A combined separation and purification device for circulating blood tumor cells

[0059] A combined sorting and purifying device for circulating blood tumor cells, including a sorting structure, a purifying structure, and a control structure 3; wherein, the control structure 3 extends out a wire to connect the sorting structure and the purifying structure; the control structure 3 controls the sorting structure and the purifying structure structure;

[0060] The sorting structure includes a microfluidic chip 11, a double-jet air pump, a nitrogen tank, and a microscope. All parts are connected to build an automatic air pump pressure-driven microfluidic chip 11 system; the overall size of the chip is 25.4*76.2mm. It includes a bottom layer 111 and a chip layer 112. On the chip layer 112, one to four column array channels 12 and two outlets and two inlets are arranged. A single array channel 12 is 68mm long, 3.5mm wide, and 50um high. The two sides of the array ch...

Embodiment 2

[0064] Example 2 Preparation method of a combined separation and purification device for circulating blood tumor cells

[0065] Preparation method of a combined separation and purification device for circulating blood tumor cells

[0066] Preparation of sorting constructs:

[0067] 1) Preparation of the microfluidic chip 11: design the structure of the regular triangular microarray column 13, and use PDMS material and soft lithography technology to make the chip; make a mask according to the designed pattern; choose polydimethylsilane ( PDMS) material; mix PDMS and curing agent according to the ratio of 1:10, vacuum air bubbles, pour into the template to vacuum air bubbles, bake in an oven at 80°C for 1 hour, take it out, cut and punch holes according to the chip design size . Take the cleaned glass slides and blow them with nitrogen, perform plasma cleaning and bonding with the cut PDMS, and bake in an oven at 80°C for 30 minutes.

[0068] 2) Connect the double-jet air pum...

Embodiment 3

[0073] Example 3 A sorting method for circulating blood tumor cells

[0074] Using the combined detection device for circulating tumor cells in Example 1 to analyze blood samples with tumor cells, specifically:

[0075] Firstly, take peripheral blood from healthy people and add traced tumor cells to form samples, the concentrations are 101 / ml, 102 / ml, 103 / ml, 104 / ml, 105 / ml to calculate the sorting rate and sorting purity of the whole device.

[0076] Pass the blood sample through the total sorting cell inlet 16 of the DLD chip, and at the same time add PBS to the total buffer inlet 17 of the chip, and then control the air pressure pump through the control structure 3 to achieve the purpose of controlling the flow rate of the liquid. The pressure pump is OB1PressureController (ELVEFLOW, France) . The flow rate of the blood sample is 1ml / min. At the collection port, almost all the red blood cells and 90% of the white blood cells flow out from the total waste liquid outlet 18, ...

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Abstract

The invention relates to a combined sorting purification device for circulating blood tumor cells. The device comprises a sorting structure, a purifying structure and a control structure, wherein guide wires of the control structure extend to connect the sorting structure and the purifying structure; the control structure controls the sorting structure and the purifying structure; the sorting structure comprises a micro-fluidic chip which comprises triangular micro-array columns on two sides and a wide channel between the micro-arrays; the purifying structure comprises a purifying chip, a permanent magnet, two stepping motors for supporting the permanent magnet to move and supporting pieces for supporting parts, the permanent magnet is arranged below the purifying chip, the two stepping motors are vertically arranged, and the permanent magnet is arranged on one of the stepping motors; the micro-fluidic chip is connected to the purifying chip through a tubular structure; the control structure comprises a power supply and a control unit; and the control structure controls the stepping motors to move. The device can sort and purify tumor cells effectively, and is simpler and more convenient.

Description

technical field [0001] The invention belongs to an apparatus for cell sorting and a cell sorting method thereof, in particular to a sorting apparatus and a sorting method for tumor cells in circulating blood, in particular to a combined sorting and purifying device for circulating blood tumor cells and a sorting and detecting device method. Background technique [0002] Tumor metastasis is one of the important reasons for the death of cancer patients. Studies have found that tumor cells (CTCs) in the circulating blood of cancer patients are closely related to patient metastasis and prognosis. The sorting of CTC is helpful to predict and judge the risk of tumor metastasis, provide important prognostic information and individualized treatment for the disease. Therefore, removing blood cells from circulating blood and isolating high-purity CTCs has become the key to diagnosing tumors and judging prognosis. However, due to the extremely small number of CTCs in peripheral blood...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/42C12M1/00C12N5/09
CPCC12N5/0693C12N2509/00
Inventor 王琪姜佳宁赵辉宋永欣陈艳田静杜小慧舒伟良
Owner 北京方远智汇科技有限公司
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