Melanocyte immunohistochemical hematoxylin-toluidine blue double staining

A technology of toluidine blue and immunohistochemistry, applied in the field of detection, to achieve the effect of simple and convenient steps, easy visual distinction, and obvious color difference

Inactive Publication Date: 2018-02-16
CHUGOKU IGAKU KAGAKUIN HIFUBIYOU KENKYUSHO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Toluidine blue stock solution is more simple and convenient to prepare than Giemsa stock solution

Method used

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  • Melanocyte immunohistochemical hematoxylin-toluidine blue double staining
  • Melanocyte immunohistochemical hematoxylin-toluidine blue double staining
  • Melanocyte immunohistochemical hematoxylin-toluidine blue double staining

Examples

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Embodiment 1

[0074] Example 1, the method of hematoxylin-toluidine blue double staining of melanin tissue immunohistochemistry, the steps are as follows:

[0075] ⑴. Routine xylene dewaxing of slices, gradient alcohol dehydration:

[0076] Xylene I and II each for 30 minutes, absolute ethanol, 95%, 90%, and 85% gradient alcohol for 3 minutes each, and washed with tap water.

[0077] ⑵. Block and inactivate endogenous peroxidase:

[0078] 3%H 2 o 2 Incubate at 37°C for 10 min, wash with PBS buffer for 5 min x 3 times;

[0079] ⑶. Antigen retrieval (routine operation);

[0080] ⑷. Serum blocking: Incubate goat serum at 37°C for 10 minutes, pour it out and do not wash;

[0081] ⑸. Add the primary antibody dropwise, PBS instead of the primary antibody as a negative control, and incubate overnight at 4°C;

[0082] ⑹. Wash with PBS buffer for 5min×3 times;

[0083] ⑺. Add the secondary antibody dropwise and incubate at room temperature for 20 minutes in the dark;

[0084] ⑻. Wash with P...

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Abstract

Hematoxylin-toluidine blue double staining method of melanocyte immunohistochemistry: ⑴. Routine xylene dewaxing of sections, gradient alcohol dehydration: ⑵. Blocking and inactivation of endogenous peroxidase: ⑶. Antigen retrieval; ⑷. Serum blocking; ⑸. Dropping primary antibody; ⑹. Dropping secondary antibody; ⑺. DAB color development; ⑻. Hematoxylin counterstaining, hydrochloric acid alcohol differentiation; ⑼. Toluidine blue working solution counterstaining, glacial acetic acid differentiation; ⑽ .Seal the slides after dehydration and transparency. The positive particles of the present invention are still brownish-yellow after restaining, and the original melanin particles in the tissue become dark green, which are distinct and easy to distinguish from brown particles without affecting the color development of hematoxylin. The nuclei of background cells are clearly visible, and immunohistochemistry Positive brown-yellow granules, dark green melanin granules, and blue nuclei have obvious color differences, which are easy to visually distinguish. The preparation of toluidine blue staining solution is simple and convenient.

Description

technical field [0001] The invention relates to a detection technology, in particular to a hematoxylin-toluidine blue double superstaining method for immunohistochemistry of melanocytes and their proliferative lesions. Background technique [0002] In the pathological research or clinical research of skin diseases, the research of human melanocytes and their proliferative lesions, especially malignant melanoma is one of the very active fields. The study of melanocytes with their own melanin granules must use immunohistochemical staining techniques for melanin tissue. The experimental steps of routine clinical non-melanin tissue immunohistochemical staining technique (ABC method) are as follows: [0003] ⑴. Routine xylene dewaxing of slices, gradient alcohol dehydration: [0004] Xylene I and II each for 30 minutes, absolute ethanol, 95%, 90%, and 85% gradient alcohol for 3 minutes each, and washed with tap water. [0005] ⑵. Block and inactivate endogenous peroxidase: ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/30
Inventor 姜祎群吴琼孙建方李阿梅邵雪宝
Owner CHUGOKU IGAKU KAGAKUIN HIFUBIYOU KENKYUSHO
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