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Circulating tumor stem cell detection kit based on magnetic beads and microfluidic chip

A microfluidic chip and tumor stem cell technology, which is applied in the field of circulating tumor stem cell detection kits, can solve the problems of chip integration obstacles, multiple steps, cross-contamination, etc., and achieve high sensitivity and specificity, convenient and fast operation, and easy collection Effect

Active Publication Date: 2017-02-01
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method can realize the capture of multi-marker cells, the repeated flow channel fixation makes the chip preparation process take a long time and many steps, and because the flow fixation is carried out in the same channel, it is easy to produce cross-contamination between upstream and downstream, resulting in false positives. In order to control the false positive rate, the length of each region must be increased, which hinders the further integration of the chip

Method used

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  • Circulating tumor stem cell detection kit based on magnetic beads and microfluidic chip
  • Circulating tumor stem cell detection kit based on magnetic beads and microfluidic chip
  • Circulating tumor stem cell detection kit based on magnetic beads and microfluidic chip

Examples

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Embodiment 1

[0031] 1. A circulating tumor stem cell detection kit based on magnetic beads and a microfluidic chip, consisting of the following components:

[0032] (1) Microfluidic chips, such as figure 1As shown, it is a schematic diagram of the microfluidic chip structure. The microfluidic chip microfluidic chip includes a glass chip substrate 1, and a microfluidic chip with a length a of 50 mm and a width b of 17 mm is provided on the glass chip substrate 1. control channel 2, and a soft magnetic microarray is arranged in the microfluidic control channel 2. The soft magnetic microarray is composed of nickel-iron alloy cylinders 3, and two adjacent rows of nickel-iron alloy cylinders 3 are arranged alternately. The distance c between two adjacent rows of nickel-iron alloy cylinders 3 is 100 μm, the diameter d of the nickel-iron alloy cylinders 3 is 30 μm, and the distance e between two adjacent rows of nickel-iron alloy cylinders 3 in each row is 50 μm. The shortest vertical distance...

Embodiment 2

[0048] 1. A circulating tumor stem cell detection kit based on magnetic beads and a microfluidic chip, the composition of which is the same as in Example 1.

[0049] 2. The preparation of the microfluidic chip is the same as in Example 1.

[0050] 3. Prepare blood samples mixed with Hep3B cell line to simulate blood samples containing circulating tumor stem cells:

[0051] (1) Hep3B cell line (purchased from Shanghai Life Science Park, Chinese Academy of Sciences, hepatocellular carcinoma tumor cells, the positive rates of Hep3B cells expressing EpCAM, CD133, CD90 and CD24 were 99.8%, 98%, 0.2% and 1%, respectively. In the embodiment, Hep3B cells are used as circulating tumor cells in the incorporation method). Take 4×10 7 Hep3B cells, 1×10 per copy 7 The cells were mixed with the following fluorescent antibodies at a ratio of 1:100: CD24-PE (Catalog no.130-095-403, Milteny Biotec), CD90-FITC (Catalog no.130-095-953, Milteny Biotec), EpCAM-PE (Catalog no.130-091-253, Milte...

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Abstract

The invention provides a circulating tumor stem cell detection kit based on magnetic beads and a microfluidic chip. The circulating tumor stem cell detection kit is characterized by comprising a microfluidic chip, at least one immunomagnetic bead, and at least one fluorescent antibody against a tumor stem cell biomarker, wherein the immunomagnetic bead can be specifically bonded with the circulating tumor stem cell and marked with the tumor stem cell biomarker; the microfluidic chip comprises a glass chip base; a microfluidic channel is arranged on the glass chip base; a soft magnetic micro-array is arranged inside the microfluidic channel. By adopting the circulating tumor stem cell detection kit, a plurality of different categories of rare circulating tumor stem cells can be captured and detected by sampling once, and the circulating tumor stem cell detection kit has high sensitivity and specificity, is convenient and fast to operate, can easily collect captured cells, and does not need complicated surface modification processes of a first antibody and a second antibody of the microfluidic channel inside the chip.

Description

technical field [0001] The invention relates to a detection kit for circulating tumor stem cells based on magnetic beads and a microfluidic chip. Background technique [0002] Hepatocellular carcinoma (HCC) (referred to as liver cancer) is one of the most common malignant tumors in the world. In China, newly diagnosed liver cancer accounts for about 55% of the world each year, and its mortality rate ranks second among malignant tumors in my country. . Although surgical resection is currently the first choice for the treatment of liver cancer, 60-70% of patients still have metastatic recurrence within 5 years after radical resection. Postoperative metastasis and recurrence of liver cancer is the main cause of death of patients after surgery, and it has become the most important obstacle to further improve the treatment effect of liver cancer. Therefore, exploring the mechanism of liver cancer metastasis and recurrence, and finding more effective means of early warning of met...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/533
CPCG01N33/57438G01N35/00G01N2035/00158
Inventor 樊嘉杨欣荣孙云帆徐泱周俭
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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