Cytochemical staining method being compatible with immunofluorescence analysis and uses thereof

A staining method and cytochemical technology, applied in the field of identification of epithelial tissue-derived tumor cells, can solve the problems of difficult morphological identification, inability to identify epithelial tissue-derived tumor cells, loss of histological relationship, etc.

Active Publication Date: 2008-07-23
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0016] Existing techniques for antigen analysis of epithelial tissue-derived tumor cells targeting tumor cell membranes and cytoplasm cannot identify all epithelial tissue-derived tumor cells that enter the blood in the aforementioned three forms, resul...

Method used

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  • Cytochemical staining method being compatible with immunofluorescence analysis and uses thereof
  • Cytochemical staining method being compatible with immunofluorescence analysis and uses thereof
  • Cytochemical staining method being compatible with immunofluorescence analysis and uses thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0086] Example 1 Cytochemical staining compatible with immunofluorescence analysis for samples obtained from lung cancer patients

[0087] 1. Negative enrichment was performed on 7.5 ml of peripheral blood obtained from a lung cancer patient (No. ZB0717). Specifically, the CD45 immunomagnetic beads (article number 130-045-801) from the German Metienne company were selected to remove the leukocytes in the peripheral blood according to the protocol given by the manufacturer, so as to obtain isolated and enriched non-blood-derived nucleated cells;

[0088] 2. Use 40 microliters of 1×PBS solution (137mM NaCl, 2.7mM KCl, 10mM NaCl, 2 HPO 4 , 2mM KH 2 PO 4 , pH7.4) after resuspension, drop evenly on the glass slide, and place it at room temperature until the solution just evaporates;

[0089] 3. Fix with 95% ethanol for 15 minutes;

[0090] 4. Soak cell slides in distilled water for 2 minutes, replace with fresh distilled water, and wash for 2 minutes;

[0091] 5. Hematoxylin ...

Embodiment 2

[0109] Example 2 Cytochemical staining for compatible immunofluorescence analysis of samples obtained from breast cancer patients

[0110] 1. Negative enrichment was performed on 7.5 ml of peripheral blood obtained from a breast cancer patient (No. ZB0715). Specifically, the CD45 immunomagnetic beads (article number 130-045-801) from the German Metienne company were selected to remove the leukocytes in the peripheral blood according to the protocol given by the manufacturer, so as to obtain isolated and enriched non-blood-derived nucleated cells;

[0111] 2. Use 40 microliters of 1×PBS solution (137mM NaCl, 2.7mM KCl, 10mM NaCl, 2 HPO 4 , 2mMKH 2 PO 4 , pH7.4) after resuspension, drop evenly on the glass slide, and place it at room temperature until the solution just evaporates;

[0112] 3. Fix with 95% ethanol for 15 minutes;

[0113] 4. Soak cell slides in distilled water for 2 minutes, replace with fresh distilled water, and wash for 2 minutes;

[0114] 5. Hematoxylin...

Embodiment 3

[0132] Example 3. Cytochemical staining compatible with immunofluorescence analysis for samples obtained from patients with esophageal cancer

[0133] 1. Negative enrichment was performed on 7.5 ml of peripheral blood obtained from a patient with esophageal cancer (number: M592). Specifically, the CD45 immunomagnetic beads (article number 130-045-801) from the German Metienne company were selected to remove the leukocytes in the peripheral blood according to the protocol given by the manufacturer, so as to obtain isolated and enriched non-blood-derived nucleated cells;

[0134] 2. Use 40 microliters of 1×PBS solution (137mM NaCl, 2.7mM KCl, 10mM NaCl, 2 HPO 4 , 2mM KH 2 PO 4 , pH7.4) after resuspension, drop evenly on the glass slide, and place it at room temperature until the solution just evaporates;

[0135] 3. Fix with 95% ethanol for 15 minutes;

[0136] 4. Soak cell slides in distilled water for 2 minutes, replace with fresh distilled water, and wash for 2 minutes; ...

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Abstract

The invention relates to a cell chemical dyeing method compatible with immunofluorescence analysis and relative applications. The invention further relates to a method for checking epithelial tissue source tumor cell in human peripheral blood sample, which uses the cell chemical dyeing method compatiable with immunofluorescence analysis. The inventive check method can simply, quickly and accurately check the tumor cells of epithelial tissue source in blood circulation from the karyocyte separated from small human periphery blood sample.

Description

technical field [0001] The invention relates to a cytochemical staining method compatible with immunofluorescence analysis and its application. The present invention also relates to a method for identifying tumor cells derived from epithelial tissue from human peripheral blood samples, wherein a cytochemical staining method compatible with immunofluorescence analysis is used. The identification method of the invention can simply, quickly and accurately identify tumor cells derived from epithelial tissue present in blood circulation from nucleated cells isolated from a small amount of human peripheral blood samples. Background technique [0002] The main methods of cytochemical staining techniques commonly used at present include: Wright staining, Giempi staining, Wright-Giemsa composite staining, Papanicolaou staining, and hematoxylin-eosin staining. The related staining techniques are summarized as follows: [0003] Wright's staining: Wright's dye contains two dyes, methyl...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N33/574C12Q1/02C12Q1/68
Inventor 赵晓航何平任传利郑朝旭赵平王成峰周兰萍许杨乔媛媛
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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