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Preparation method of cell staining quality control product

A quality control product and cell technology, applied in the field of quality control product preparation for cell staining, can solve the problem of low accuracy of circulating tumor cells in lung cancer

Active Publication Date: 2021-06-29
深圳天烁生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The main purpose of the present invention is to propose a quality control product preparation method for cell staining, aiming to solve the technical problem that the accuracy of the existing staining kits for detecting circulating tumor cells in lung cancer is not high

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  • Preparation method of cell staining quality control product
  • Preparation method of cell staining quality control product
  • Preparation method of cell staining quality control product

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preparation example Construction

[0032] The present invention proposes a method for preparing a quality control product for cell staining, comprising the following steps: (1) E-Cadherin immunomagnetic beads are mixed with a binding buffer to obtain E-Cadherin immunomagnetic beads working solution, and CEA immunomagnetic beads are mixed with The binding buffer is mixed to obtain the CEA immunomagnetic beads working solution, and the CD45 immunomagnetic beads are mixed with the binding buffer to obtain the CD45 immunomagnetic beads working solution; (2) the E-Cadherin immunomagnetic beads working solution, the CEA Mixing the immunomagnetic bead working solution, the CD45 immunomagnetic bead working solution, leukocytes and lung cancer circulating tumor cells to obtain the first mixed solution; (3) removing the liquid in the first mixed solution, adding a fixative to fix it, and obtaining Quality control of circulating tumor cells in lung cancer.

[0033] In the preparation method of the present invention, E-cad...

Embodiment 1

[0043] 1. Mix white blood cells with NCI-H1993 cells (number ratio: 1:10), the total volume is 700ul;

[0044] 2. Take 30ul of E-Cadherin immunomagnetic beads, add 1ml of binding buffer, discard the supernatant, and resuspend the magnetic beads with 1ml of binding buffer to make E-Cadherin immunomagnetic beads working solution;

[0045] 3. Treat CEA immunomagnetic beads and CD45 immunomagnetic beads with the same method as step 2 to make CEA immunomagnetic beads working solution and CD45 immunomagnetic beads working solution;

[0046] 4. Add 30ul of the E-Cadherin immunomagnetic bead working solution, 40ul of the CEA immunomagnetic bead working solution, and 50ul of the CD45 immunomagnetic bead working solution into the cell mixture in step 1, and mix evenly by inversion. Discard the supernatant and repeat the operation 3 times;

[0047] 5. Rotate and mix the centrifuge tube in a refrigerator at 4°C for 2 hours at a rotation speed of 15 rpm;

[0048] 6. After rotating and mi...

Embodiment 2

[0056] 1. Mix white blood cells with NCI-H1993 cells (the ratio of cell number is 10:1), and the total volume is 700ul;

[0057] 2. Take 30ul of E-Cadherin immunomagnetic beads, add 1ml of binding buffer, discard the supernatant, and resuspend the magnetic beads with 1ml of binding buffer to make E-Cadherin immunomagnetic beads working solution;

[0058] 3. Treat CEA immunomagnetic beads and CD45 immunomagnetic beads with the same method as step 2 to make CEA immunomagnetic beads working solution and CD45 immunomagnetic beads working solution;

[0059] 4. Add 40ul of the E-Cadherin immunomagnetic bead working solution, 30ul of the CEA immunomagnetic bead working solution, and 28ul of the CD45 immunomagnetic bead working solution into the cell mixture in step 1, and mix evenly by inversion. Discard the supernatant and repeat the operation 3 times;

[0060] 5. Rotate and mix the centrifuge tube in a refrigerator at 4°C for 2 hours at a rotation speed of 15 rpm;

[0061] 6. Aft...

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Abstract

The invention discloses a preparation method of a cell staining quality control product. According to the preparation method, E-Cadherin immunomagnetic beads and CEA immunomagnetic beads are specifically combined with lung cancer circulating tumor cells, and CD45 immunomagnetic beads are specifically combined with white blood cells; by staining the enriched lung cancer circulating tumor cells and leukocytes, the method can conduct quality control on a staining kit for the lung cancer circulating tumor cells. According to the preparation method of the cell staining quality control product, through specific binding of the antigen and the antibody, the credibility of a quality control result is high.

Description

technical field [0001] The invention relates to the field of cell staining detection, in particular to a method for preparing a quality control product for cell staining. Background technique [0002] Cancer cells shed from the primary site and enter the peripheral blood of the human body, which are called circulating tumor cells (Circulating Tumor Cell, CTC). Lung cancer mainly originates from the mucosal epithelium of the bronchi, and has become the cancer with the highest incidence and the largest number of deaths in the world. In lung cancer patients, circulating tumor cells of lung cancer metastasize to other organs of the body through blood dissemination, which is the main cause of death of patients. Therefore, detection of changes in the content of circulating tumor cells of lung cancer in peripheral blood is of positive significance for real-time monitoring of the dynamics of lung cancer cells in patients and evaluation of therapeutic effects. [0003] However, the...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N33/68G01N33/574
CPCG01N1/30G01N33/57423G01N33/57473G01N33/57488G01N33/6872G01N33/56966G01N2333/70589
Inventor 张磊
Owner 深圳天烁生物科技有限公司
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