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Cast-off cell one-step staining method, and dye combination and kit for cast-off cell one-step staining method

A technique of exfoliated cells and dyeing method, applied in the dye combinations and kits used, in the field of exfoliated cells one-step dyeing method, which can solve the problems of easy co-staining, time-consuming, affecting accuracy, etc., achieving rapid dyeing and detection, simplifying light source, Avoid the effects of complexity

Inactive Publication Date: 2018-05-08
陈石磊
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] (1) The dyeing operation process is complicated, the liquid preparation and reaction conditions are strict, and it takes a long time (several hours)
[0009] (2) The specificity of staining is not strong, and the counterstained HE or EA36 background is easy to co-stain, which directly affects the accuracy of interpretation
[0011] (4) After staining, cell sections are easy to fade and cannot be stored for a long time
Physicians cannot extend the accumulated (Pap stain) experience to read films
Due to the above shortcomings, Feulgen staining has not been widely used clinically

Method used

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  • Cast-off cell one-step staining method, and dye combination and kit for cast-off cell one-step staining method
  • Cast-off cell one-step staining method, and dye combination and kit for cast-off cell one-step staining method
  • Cast-off cell one-step staining method, and dye combination and kit for cast-off cell one-step staining method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0125] 1. Fluorescent dye buffer configuration

[0126] 1. 0.1M citric acid mother liquor: weigh 2.1g citric acid monohydrate, dissolve in 100ml dH 2 O middle;

[0127] 2. 0.2M disodium hydrogen phosphate: Weigh 7.16g disodium hydrogen phosphate dodecahydrate, dissolve in 100ml dH 2 O middle;

[0128] 3. 10mM citric acid-phosphate buffer, 0.1M NaCl: Take 9.92ml 0.1M citric acid, 5.46ml 0.2M disodium hydrogen phosphate, 1.7g NaCl, add water to 200ml and stir to dissolve, adjust the pH to 3.5.

[0129] The above 10mM citric acid-phosphate buffer solution, pH3.5, 0.1M NaCl is the fluorescent dye preservation solution.

[0130] 2. Fluorescent dye working solution configuration

[0131] 1. Accurately weigh 5mg of acridine orange (AO), dissolve it in 10ml of 10mM citric acid-phosphate buffer, use it as acridine orange (AO) mother solution (0.5mg / ml), store at 2-8°C;

[0132] 2. Accurately weigh 0.25mg of DAPI, dissolve in 10mldH 2 O, as DAPI stock solution (0.025mg / ml), stored...

Embodiment 2

[0148] 1. Fluorescent dye buffer configuration

[0149] 1. 0.1M citric acid mother liquor: weigh 2.1g citric acid monohydrate, dissolve in 100ml dH 2 O middle;

[0150] 2. 0.2M disodium hydrogen phosphate: Weigh 7.16g disodium hydrogen phosphate dodecahydrate, dissolve in 100ml dH 2 O middle;

[0151] 3. 10mM citric acid-phosphate buffer, 0.1M NaCl: Take 9.92ml 0.1M citric acid, 5.46ml 0.2M disodium hydrogen phosphate, 1.7g NaCl, add water to 200ml and stir to dissolve, adjust the pH to 2.

[0152] The above 10mM citric acid-phosphate buffer solution, pH 2, 0.1M NaCl is the fluorescent dye preservation solution.

[0153] 2. Fluorescent dye working solution configuration

[0154] 1. Accurately weigh 100mg of acridine orange (AO), dissolve it in 10ml of 10mM citric acid-phosphate buffer, use it as acridine orange (AO) mother solution (10mg / ml), store at 2-8°C;

[0155] 2. Accurately weigh 0.25mg of DAPI, dissolve in 10mldH 2 O, as DAPI stock solution (0.025mg / ml), stored at ...

Embodiment 3

[0171] 1. Fluorescent dye buffer configuration

[0172] 1. 0.1M citric acid mother liquor: weigh 2.1g citric acid monohydrate, dissolve in 100ml dH 2 O middle;

[0173] 2. 0.2M disodium hydrogen phosphate: Weigh 7.16g disodium hydrogen phosphate dodecahydrate, dissolve in 100ml dH 2 O middle;

[0174] 3. 10mM citric acid-phosphate buffer, 0.1M NaCl: Take 9.92ml 0.1M citric acid, 5.46ml 0.2M disodium hydrogen phosphate, 1.7g NaCl, add water to 200ml and stir to dissolve, adjust the pH to 6.

[0175] The above 10mM citric acid-phosphate buffer solution, pH 6, 0.1M NaCl is the fluorescent dye preservation solution.

[0176] 2. Fluorescent dye working solution configuration

[0177] 1. Accurately weigh 1mg of acridine orange (AO), dissolve it in 10ml of 10mM citric acid-phosphate buffer, use it as acridine orange (AO) mother solution (0.1mg / ml), store at 2-8°C;

[0178] 2. Accurately weigh 1.25mg of DAPI, dissolve in 10mldH 2 In O, as DAPI mother solution (0.125mg / ml), store...

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Abstract

The invention belongs to the field of cytopathology, and particularly relates to a cast-off cell one-step staining method, and a dye combination and a kit for the cast-off cell one-step staining method, wherein one-step staining is the synchronous staining of the nucleus and the cytoplasm of cast-off cells, and comprises: carrying out one-step staining on cast-off cells by using a fluorescent dyecombination, and the fluorescent dye combination contains a nucleus fluorescent dye and a cytoplasm fluorescent dye. According to the present invention, the nucleus and the cytoplasm of cast-off cellsare synchronously stained with the fluorescent dye combination without the washing of the background fluorescence, such that the rapid staining and detection can be achieved; and excitation is performed with the single ultraviolet light source, and the good effect is achieved, such that the light source can be simplified, and the image forming complexity caused by the two irradiations can be avoided.

Description

technical field [0001] The invention belongs to the field of cytopathology, and in particular relates to a one-step staining method for exfoliated cells, a used dye combination and a kit. Background technique [0002] Exfoliative cytology examination refers to the collection of exfoliated epithelial cells on the surface of human luminal organs, and after staining, observe their morphology with a microscope and make a diagnosis. It is widely used in precancerous lesion screening. The main sources of exfoliated cells are: cervical exfoliated cells, serous cavity effusion exfoliated cells and urinary system exfoliated cells. [0003] The conventional exfoliated cell staining method uses the different biochemical compositions of various structures in the cells to react with eosinophilic or basophilic dyes to display different colors, making the shape and structure of cells easy to identify. Commonly used are Papanicolaou and Wright-Geek staining. [0004] Papanicolaou stainin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
CPCG01N1/30G01N2001/302
Inventor 陈石磊赵路赵志炜
Owner 陈石磊
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