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Application of Shorr staining reagent and staining method

A dyeing reagent and dyeing method technology, applied in the field of Shorr dyeing reagents

Pending Publication Date: 2021-10-08
THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it takes 2-3 working days for the routine pathological diagnosis report of testicular biopsy to be issued, which cannot provide a reliable diagnostic basis for clinical testicular sperm retrieval in real time

Method used

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  • Application of Shorr staining reagent and staining method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The method for staining sperm provided in this embodiment comprises the following steps:

[0032] 1 Frozen section preparation: The temperature of the frozen microtome is -18°C to -22°C. The fresh testicular biopsy tissue was embedded with OCT cryo-embedding gel and then frozen for about 1 min, and sliced ​​into 4 μm sections by a cryostat to obtain frozen sections of the testicular biopsy tissue for future use.

[0033] 2. Shorr staining step (the kit was purchased from Zhuhai Beisuo Biotechnology Co., Ltd., Yuezhu Machinery No. 20160013): After the frozen section prepared in step 1 was made, it was naturally dried in the air, fixed with a fixative for 30 seconds to 1 minute, and flowed slowly Rinse; (2) stain with hematoxylin solution for 1-2 minutes, rinse slowly with running water; (3) immerse frozen sections in acidic ethanol for differentiation for 8-10 seconds, and rinse with running water for at least 2 minutes (in other embodiments, to save time, immerse in sat...

experiment example 1

[0041]Collected 30 testicular biopsy specimens from the Andrology Department of the Third Affiliated Hospital of Guangzhou Medical University from February 2019 to April 2019. All specimens were fresh tissue punctured during the operation. The specimens were sent to the pathology department for freezing immediately after isolation. Sections were stained by conventional HE staining (Comparative Example 1), standard Papanicolaou staining (Comparative Example 1), Diff-Quik staining (Comparative Example 1) and Shorr staining (Example 1). Under the optical microscope, two pathologists jointly evaluated the effects of four kinds of histochemical staining on the frozen sections of the testicular biopsies of 30 patients with OA, and evaluated the sperm morphology, staining effect and background cell contrast. The results are shown in Table 1 and figure 1 shown.

[0042] Table 1 Comparison of the color, contrast and staining time of each part of sperm by 4 staining methods

[0043] ...

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Abstract

The invention discloses application of a Shorr staining reagent and a staining method, and relates to the technical field of assisted reproduction. The invention discloses application of a Shorr staining reagent in preparation of a sperm morphology diagnostic kit. A sample of the diagnostic kit is a testis needle biopsy tissue frozen section. According to the kit, fresh testis needle biopsy tissue frozen sections serve as staining objects, all parts of sperms can be stained in different colors, the sperms and background cells can be effectively distinguished, the recognition degree of the sperms is improved, and the diagnosis time of the pathological morphology of the sperms is shortened.

Description

technical field [0001] The invention relates to the technical field of assisted reproduction, in particular to the application of Shorr staining reagent and a staining method. Background technique [0002] With the development of assisted reproductive technology, especially the intracytoplasmic sperm injection technology, a breakthrough has been made in the treatment of patients with azoospermia. This technique uses micromanipulation techniques to inject sperm into the egg cell cytoplasm to fertilize the egg. Clinically successful sperm extraction, especially high-quality sperm is the prerequisite for single sperm injection treatment. Testicular sperm aspiration is the main method for obtaining sperm in patients with azoospermia. The histopathological diagnosis of testicular biopsy can directly observe the state of spermatogenesis and the degree of damage in the seminiferous tubules, and can accurately evaluate the spermatogenesis state and whether there are sperm in the t...

Claims

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Application Information

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IPC IPC(8): G01N1/31
CPCG01N1/31
Inventor 陈辉
Owner THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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