Sperm fluorescent dyeing determination method

A fluorescent dyeing and measuring method technology, applied in biological testing, material inspection products, etc., can solve the problems of sperm separation, poor repeatability, inability to distinguish immobile sperm and dead sperm, etc.

Inactive Publication Date: 2008-11-19
SHANGHAI INST OF PLANNED PARENTHOOD RES
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AI Technical Summary

Problems solved by technology

The use of computer-assisted sperm analysis (CASA) can provide more accurate quantitative data on sperm dynamics, but the current clinical use of CASA cannot be used for the detection of sperm density because it is difficult to distinguish sperm from non-sperm granular debris
Another disadvantage of the CASA system is that it cannot distinguish between motile and dead sperm, especially in male patients requiring assisted reproductive therapy. If the percentage of motile sperm (grade D) exceeds 50%, the sperm viability should be tested, and Viability assessment verifies accuracy of sperm motility assessment
[0003] The most commonly used tests for sperm viability are eosin staining and sperm tail hypotonic swelling test. The biggest disadvantage of the two methods is poor repeatability, the results of different laboratories vary greatly, and the operation process has an impact on sperm viability. Significant effects, some sperm are even immobile

Method used

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  • Sperm fluorescent dyeing determination method
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Experimental program
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Embodiment 1

[0060] Sperm fluorescent staining

[0061] Materials and Methods

[0062] Research object:

[0063] Normal fertile male with sperm density >20×10 6 / ml, grade a+b sperm>50%, WBC6 / ml, after 3-5 days of abstinence, collect the semen in a clean glass container by masturbation. After the semen is liquefied, the spermatozoa are optimized by the upstream method and adjusted to a density of 60×10 6 / ml, the total activity rate>90%.

[0064] Reagent:

[0065] Propidium iodide (PI) was purchased from Sigma Company;

[0066] Transgreen fluorescent molecular probes were purchased from Shanghai Ganfeng Biotechnology Co., Ltd.

[0067] instrument:

[0068] Computer-aided semen analysis system (with fluorescence device and constant temperature device), VideoTest Cytogenetics automatic sperm image analysis system provided by Anlai Company;

[0069] CO 2 Incubator, purchased from Heraeus.

[0070] method:

[0071] 1. Sperm motility analysis

[0072] Divide the processed sperm sus...

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Abstract

The invention discloses a sperm fluorescence staining method, which comprises the steps of: mixing sperms with PI, Transgreen and Transgreen / PI, and putting the mixture into an incubator with 5 percent CO2 and temperature of 37 DEG C for cultivation. Applying a computer to assist a sperm and semen quality analysis system and analyze changes of all movement parameters of sperms after adding fluorescent dye and Transgreen / PI redyeing is feasible for calculating the survival rate of sperms.

Description

technical field [0001] The invention relates to fluorescent staining, in particular to nucleic acid staining, and more particularly to sperm fluorescent staining. Background technique [0002] Sperm density, sperm membrane integrity, and sperm motility are the most commonly used indicators for evaluating semen quality in clinical andrology laboratories. The use of computer-assisted sperm analysis (CASA) can provide more accurate quantitative data on sperm dynamics, but the current clinical use of CASA cannot be used for the detection of sperm density because it is difficult to distinguish sperm from non-sperm granular debris . Another disadvantage of the CASA system is that it cannot distinguish between motile and dead sperm, especially in male patients requiring assisted reproductive therapy. If the percentage of motile sperm (grade D) exceeds 50%, the sperm viability should be tested, and The viability assessment verifies the accuracy of the sperm motility assessment. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/48
Inventor 施惠娟王健袁瑶赵洪鑫
Owner SHANGHAI INST OF PLANNED PARENTHOOD RES
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