Breast cancer cell dyeing method, application thereof and dyeing kit

A technique for breast cancer cells and a staining method, which is applied in the field of breast cancer cell staining methods and applications and staining kits, can solve problems such as poor prognosis, and achieve the effects of good application prospects, simple operation and good accuracy.

Inactive Publication Date: 2016-01-06
JIANGSU PROVINCE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous studies have found that micrometastasis detection can reflect the distant metastasis of patients earlier than current follow-up...

Method used

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  • Breast cancer cell dyeing method, application thereof and dyeing kit
  • Breast cancer cell dyeing method, application thereof and dyeing kit
  • Breast cancer cell dyeing method, application thereof and dyeing kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Staining of breast cancer cell lines.

[0049] (1) First prepare a glass slide on which the breast cancer cell line MCF-7 is tiled on the surface: digest the breast cancer cell line MCF-7 cultured in vitro, add 1 ml of PBS solution, dilute and mix well.

[0050] (2) Turn on the Hettich centrifuge, add samples with the PBS containing breast cancer cell line MCF-7 in step (1), so that the total cell volume of each sample well is 0.3M, and discard the PBS solution after centrifugation to obtain a surface The slides of breast cancer cell MCF-7 were tiled, air-dried, and fixed with acetone for 10 min.

[0051] (3) Use an immunohistochemical pen to draw a circle around the tissue of the slide obtained in step (2), the circle is about 3mm away from the tissue, wash with PBS for 3min, and then wash with 3wt%H 2 o 2 Soak at room temperature for 10 minutes, and then wash with PBS 3 times, 3 minutes each time, to obtain a glass slide with breast cancer cells spread on ...

Embodiment 2

[0056] Example 2 Staining detection of breast cancer cell line MCF-7 added to normal peripheral blood samples.

[0057] 1) Collect 5ml of peripheral blood sample with a sodium heparin anticoagulant tube, add a small amount of PBS solution containing breast cancer cell line MCF-7, and mix well.

[0058] 2) Centrifuge in a centrifuge (3500 rpm, 5 minutes), absorb and discard the upper layer of plasma, and obtain about 2-3ml of blood cells.

[0059] 3) After diluting and mixing the remaining blood cell components with 3ml PBS, add 15ml Ficoll (lymphocyte separation medium, Lymphoprep, Axis-Shield PoCAS, Oslo, Norway) into a centrifuge tube, and centrifuge for 20min in a horizontal centrifuge (22°C, 2000 rpm, 0 speed to start, 0 speed to decelerate to stop).

[0060] 4) Take out the specimen from the centrifuge, extract the mononuclear cell layer cells, add to the PBS solution and mix well, centrifuge (2000 rpm, 10min), discard the original PBS, and add 2ml of new PBS to dilute a...

Embodiment 3

[0063] Example 3 Staining detection by adding breast cancer cell lines to bone marrow blood samples.

[0064] 1) Collect 5ml of bone marrow blood sample with a sodium heparin anticoagulant tube, add a small amount of PBS solution containing breast cancer cell line MCF-7, and mix well.

[0065] 2) Centrifuge in a centrifuge (3500 rpm, 5 minutes), absorb and discard the upper layer of plasma, and obtain about 2-3ml of blood cells.

[0066] 3) After diluting and mixing the remaining blood cell components with 3ml PBS, add 15ml Ficoll (lymphocyte separation medium, Lymphoprep, Axis-Shield PoCAS, Oslo, Norway) into a centrifuge tube, and centrifuge for 20min in a horizontal centrifuge (22°C, 2000 rpm, 0 speed to start, 0 speed to decelerate to stop).

[0067] 4) Take out the specimen from the centrifuge, extract the mononuclear cell layer cells, add to the PBS solution to mix, centrifuge (2000 rpm, 10min), discard the original PBS, add 2ml of new PBS to dilute and mix, spare.

...

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Abstract

The invention provides a breast cancer cell dyeing method. The method takes anti-pan cytokeratin antibody (AE1/AE3) as the primary antibody, and adopts a horseradish peroxidase labeled rabbit antimouse antibody as the second antibody to label breast cancer cells, by means of diaminobenzidine (DAB) and hematoxylin dyeing, breast cancer cells can be dyed specifically. The invention also puts forward application of the dyeing method and a kit for carrying out the method. The method provided by the invention has the advantages of simple operation and good accuracy, can be used for detection of breast cancer micrometastatic cells in bone marrow blood and peripheral blood samples of breast cancer patients, and has important reference significance to prognosis of breast cancer.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a breast cancer cell staining method, its application and a staining kit. Background technique [0002] The long-term survival evaluation indicators of breast cancer patients include the size of the primary tumor, the presence or absence of axillary lymph node metastasis, tumor molecular classification, and evidence of distant metastasis. These indicators are very meaningful for the survival evaluation of patient groups. However, the accuracy of risk judgment for some individual patients is relatively limited. In addition, during the follow-up period of breast cancer, regular physical examinations are carried out through physical examinations, imaging and other examinations. By the time these examinations find problems, the best time for treatment has actually been missed. In the early stages of breast cancer, a considerable number of patients have micro...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N1/31
Inventor 王水肇毅王珏
Owner JIANGSU PROVINCE HOSPITAL
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