Supercharge Your Innovation With Domain-Expert AI Agents!

A method of detecting human sperm vitality

A technology for sperm motility and human detection, applied in material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of inaccurate results, reduced sperm motility, and inaccurate test results.

Active Publication Date: 2015-09-09
SICHUAN NEO LIFE STEM CELL BIOTECH
View PDF3 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current detection of sperm motility by flow cytometry is relatively extensive and not accurate enough. The main reason is that the accuracy of flow cytometry detection results depends on the selection and analysis of detection data, especially the adjustment and compensation stage is very critical. However, at present Most technicians will not adjust the compensation or have difficulty finding the correct compensation value, resulting in inaccurate results
For example, Xia Xinyi et al., "Studies on the Detection of Sperm Mitochondrial Membrane Potential by JC-1 Single-label Flow Cytometry", Chinese Journal of Andrology, Volume 14, Issue 2, February 2008, disclosed a method based on JC-1 The dye is a method for detecting sperm motility by flow cytometry, and the flow cytometry detection does not have compensation, and the detection results are inaccurate
[0004] In addition, due to the high viscosity of semen, it is easy to coagulate. At present, trypsin or repeated centrifugation after dilution are used to solve this problem. However, these treatments will reduce the motility of sperm and affect the accuracy of test results.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A method of detecting human sperm vitality
  • A method of detecting human sperm vitality
  • A method of detecting human sperm vitality

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1 The present invention detects the method for human sperm motility

[0033] 1. Test method

[0034] Obtain semen, dilute 10 times with PBS, add JC-1 to a final concentration of 2 μg / ml, and treat at 37°C for 10 minutes; the mixture is not centrifuged, and directly analyzed by flow cytometry, and the fluorescence detection uses excitation light with a wavelength of 488nm , Collect data of FS Lin, SS Log, FL1 Log, FL2 Log and other parameters during detection.

[0035] 2. Data analysis

[0036] For flow cytometry data: take the FS Lin of all cells as the X axis, and the SS Log as the Y axis, draw a scatter diagram, and set a gate to circle the main cell population and remove cell debris. Based on this gate, the The FL1 Log of the cells inside the gate is the X axis, and the FL2 Log of the cells inside the gate is the Y axis, draw a scatter plot, and then adjust the compensation, the compensation is: FL1-8% FL2 and FL2-10% FL1 (see figure 1 ), and then set a...

experiment example 2

[0059] Verification of the semen pretreatment method in the method of the present invention in experimental example 2

[0060] 1. The semen pretreatment method of the present invention can solve the problems of non-liquefied sperm and high viscosity

[0061] In the case of non-liquefied sperm and high viscosity, the error of the detection value obtained by conventional microscopic analysis is extremely large, and it is also extremely unfavorable for flow cytometry detection.

[0062] The inventor found through exploration that diluting 10 times with PBS can solve the problems of non-liquefaction and high viscosity of sperm. The physical pictures of "normally liquefied semen" and "non-liquefied semen" are shown in Figure 4 .

[0063] Two, the comparison of the inventive method and traditional method to sperm damage

[0064] (1) Microscopic observation

[0065] The semen pretreatment of the method of the present invention: obtain semen, dilute 10 times with PBS, to be teste...

experiment example 3

[0082] The verification of setting up gate mode in the method of the present invention in experimental example 3

[0083] In the original FS / SS scatter diagram, there are discrete points relative to the main group. These points are impurity fragments. If not excluded, they will affect the analysis. Therefore, these impurity fragments can be removed by setting a gate so that the analysis can proceed normally.

[0084] The gating method of the method of the present invention is to set a gate for scattered light, specifically, according to the obviously existing clustered cell group in the scatter diagram and the scattered point groups at its edge, circle the obviously existing clustered cell group with a rectangular gate method, Scattered point groups at their edges are excluded.

[0085] The gating method of the present invention is established based on the principles of FS and SS. FS, as forward scattering, can objectively reflect the size of particles, and the particle size a...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

A method of detecting human sperm vitality is provided. A flow cytometer is adopted for detection. The method includes (1) a step of preparing a seminal fluid to be detected, and diluting with PBS by 10 times to obtain a diluted seminal fluid; (2) a step of performing cell staining by using fluorochrome JC-1; (3) a step of detecting, namely a step of detecting by using the flow cytometer with the excitation wavelength being 488 nm to obtain FS Lin, SS Log, FL1 Log and FL2 Log values of each cell; and (4) a step of data analysis, namely a step of mapping a scatter diagram by adopting the FS Lin of all the cells as an X axis and adopting the SS Log of all the cells as a Y axis, gating, mapping a scatter diagram by adopting the FL1 Log of cells in the gates as an X axis and adopting the FL2 Log of cells in the gates as a Y axis, adjusting, compensating that is FL1-8%FL2 and FL2-10%FL1, gating, and counting cells in the gates. The method can accurately detect sperm vitality and is simple, convenient and rapid.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a method for detecting human sperm viability. Background technique [0002] Sperm quality or sperm motility is an important criterion for evaluating male fertility. Accurate and objective evaluation of sperm quality is the key to the success of artificial insemination and in vitro fertilization techniques. The indicators and evaluation methods used to reflect sperm quality include: sperm morphology, survival rate , activity rate, acrosomal membrane integrity, sperm mitochondrial activity, etc. At present, the gold standard for detecting sperm motility is direct observation with a microscope, but microscope observation has the problems of time-consuming, laborious, and low detection efficiency, and it is difficult to meet the needs of clinical detection. [0003] Flow cytometry can quickly analyze the state of cells with high throughput, and can detect multiple fluorescence pa...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 万谦陈强陆华高小平
Owner SICHUAN NEO LIFE STEM CELL BIOTECH
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com