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Sperm morphology rapid dyeing reagent and dyeing method thereof

A technology of rapid dyeing and dyeing method, applied in the field of sperm morphology rapid dyeing reagent and its dyeing, can solve the problems of high cost and complicated sperm dyeing operation, and achieve the effects of low cost, abundant raw material sources and long storage period

Inactive Publication Date: 2019-03-01
山东康华生物医疗科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the disadvantages of complicated sperm staining operation and high cost in the prior art, and propose a rapid staining reagent for sperm morphology

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0032] Preparation of phosphate buffer: prepare 1000 ml of 0.2M potassium dihydrogen phosphate and 1000 ml of 0.2M disodium hydrogen phosphate, mix the above two solutions under the monitoring of a pH meter, and adjust the pH 7.4 phosphate buffer 1: After 10 dilution, add 0.1% sodium azide.

[0033] Preparation of solution A: accurately weigh 0.001g-0.002g of Fast Green, add methanol to 1000ml, fully dissolve it, filter and store at room temperature.

[0034] Preparation of solution B: accurately weigh out 0.8g-1.22g of eosin, add 10ml sodium azide, add phosphate buffer to 1000ml, fully dissolve it, filter and store at room temperature.

[0035] Preparation of Solution C: Weigh 1.625g-2g of basic methylene blue and 1.625g-2g of methylene blue, add PB buffer to 1000ml, fully dissolve, filter and store at room temperature.

Embodiment 1

[0037] A rapid staining reagent for sperm morphology, including solution A, solution B and solution C;

[0038] Solution A is a tissue fixative containing solid green, and solution A is used to stain the acrosome area of ​​sperm, and the tissue fixative in solution A is methanol;

[0039] Solution B is an antiseptic solution containing eosin, and solution B is used to dye the sperm body and tail area, and the antiseptic solution in solution B is sodium azide solution;

[0040] Solution C is a phosphate buffer containing alkaline sky blue A and methylene blue, and solution C is used to stain the posterior acrosome area of ​​sperm.

[0041] Among them, the concentration of solid green in solution A is 0.001g / L; the concentration of eosin in solution B is 1g / L; the concentration of alkaline sky blue A in solution C is 0.1g / L, and the concentration of methylene blue is 0.1g / L .

[0042] Among them, the pH of the phosphate buffer is 7.0.

[0043] A method for staining sperm morphology rapid ...

Embodiment 2

[0051] A rapid staining reagent for sperm morphology, including solution A, solution B and solution C;

[0052] Solution A is a tissue fixative containing solid green, and solution A is used to stain the acrosome area of ​​sperm, and the tissue fixative in solution A is methanol;

[0053] Solution B is an antiseptic solution containing eosin, and solution B is used to dye the sperm body and tail area, and the antiseptic solution in solution B is sodium azide solution;

[0054] Solution C is a phosphate buffer containing alkaline sky blue A and methylene blue, and solution C is used to stain the posterior acrosome area of ​​sperm.

[0055] Among them, the concentration of solid green in solution A is 0.005 g / L; the concentration of eosin in solution B is 2 g / L; the concentration of alkaline sky blue A in solution C is 1 g / L, and the concentration of methylene blue is 1 g / L.

[0056] Among them, the pH value of the phosphate buffer is 8.0.

[0057] A method for staining sperm morphology ra...

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Abstract

The invention discloses a sperm morphology rapid dyeing reagent and a dyeing method thereof. The sperm morphology rapid dyeing reagent is prepared from a solution A, a solution B and a solution C; thesolution A is tissue fixative containing fast green; the solution B is a preservation solution containing eosin; and the solution C is phosphate buffer containing alkaline azure A and methylthioninechloride. The dyeing method comprises the steps that fresh seminal fluid is collected and liquidized in a water-bath pot with the temperature being 20-37 DEG C, sperms are optimized for 8-12 minutes by adopting a swim-up method, and the fresh sperms with high activity are obtained; sperm suspension and a smear are prepared, and the smear is dried under a sterile environment; the smear is dipped into the solution A and reacts for 15 seconds at room temperature so as to remove redundant fixative; the smear is dipped into the solution B and reacts for 10 seconds at room temperature so as to remove the redundant dyeing agent; the smear is dipped into the solution C and reacts for 10 seconds at room temperature so as to remove the redundant reagent; the redundant dyeing agent is removed; moisture is removed, and after the smear is completely dried, dyeing is completed. The sperm morphology rapid dyeing reagent well solves the defects that sperm dyeing is complex to operate, and the cost ishigh in the prior art.

Description

Technical field [0001] The invention relates to the field of human sperm detection, in particular to a rapid staining reagent for sperm morphology and a staining method thereof. Background technique [0002] With the gradual increase in the incidence of infertility, effective diagnosis and treatment of male infertility is also particularly urgent. Investigations and studies have shown that 10-15% of couples of childbearing age are infertile, of which male causes account for about 50%. Sperm is the ultimate performer of male reproduction, the end product of male germ cell line development, and the carrier of stable transmission of parental genetic information to offspring. Therefore, normal sperm is the basic guarantee for male fertility. [0003] Sperm morphology testing is an important indicator in the diagnosis of male infertility, and it plays an important role in the selection of assisted reproduction methods. [0004] At present, there are many kinds of sperm staining technique...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30G01N1/44
CPCG01N1/30G01N1/44G01N2001/302G01N2001/305
Inventor 杨致亭郭占山候鹏赵鹏潘德芹
Owner 山东康华生物医疗科技股份有限公司
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