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Methods in molecular biology (Clifton, N.J.)2025; 2954; 241-259; doi: 10.1007/978-1-0716-4698-4_14

Analysis of Motion Characteristics and Plasma Membrane Intactness (Viability) in Sperm from Domestic Animals.

Abstract: Sperm quality analysis using computer-assisted sperm analysis (CASA) systems and fluorescence-based techniques has become common in the animal reproduction industry, particularly for large animals (i.e., bovine, porcine, equine). In this chapter, the methods commonly utilized in the author's laboratory to examine sperm motion characteristics via CASA and plasma membrane intactness by flow cytometry will be described. These include methods to properly dilute fresh (stallions, bulls, boars), cool-stored (stallions, boars), or frozen/thawed (stallions, bulls, boars) sperm for assessment of sperm motion characteristics using CASA, preparation of fluorochromes (SYBR-14/propidium iodide) utilized to distinguish between intact (i.e., viable or "live") versus non-intact (i.e., nonviable or "dead") sperm, and the analysis and interpretation of plasma membrane intactness using flow cytometry.
© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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Publication Date: 2025-07-02 PubMed ID: 40601280DOI: 10.1007/978-1-0716-4698-4_14Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article illustrates various methods for analysing the quality of animal sperm, specifically focusing on techniques used in assessing motion characteristics of sperm and their plasma membrane intactness (viability).

Article Explanation

The featured article’s overarching focus is an analysis of sperm quality in large domestic animals, including bovine (cows), porcine (pigs), and equine (horses). The techniques outlined are designed to evaluate the sperm’s motion characteristics and the intactness of the plasma membrane, which is a critical measure of sperm viability.

  • Methods Used: The primary methods used in the author’s laboratory are discussed in the paper. They include the Computer-Assisted Sperm Analysis (CASA) systems and fluorescence-based flow cytometry. CASA helps in the examination of the sperm’s motion characteristics, whilst flow cytometry is employed to evaluate the sperm’s plasma membrane intactness. The plasma membrane of a sperm must be intact for the sperm to be considered viable.
  • Sperm Processing: To prepare the sperm for analysis, it is first diluted. The procedures for dilution could alter depending on whether the sperm under study is fresh (from stallions, bulls, and boars), cool-stored (from stallions and boars), or has been frozen and thawed (from stallions, bulls and boars), and such specifics are explained in detail in the research piece.
  • Fluorochromes Preparation: Fluorochromes, specifically SYBR-14 and propidium iodide, are prepared for use in distinguishing between ‘live’ (intact) sperm and ‘dead’ (non-intact) sperm. The article goes into the preparation of these vital distinguishing agents.
  • Analysis and Interpretation: After the process of sperm dilution and fluorochromes preparation, analysis and interpretation are conducted using flow cytometry. This analysis process determines the intactness of the plasma membrane, thus providing vital information about the sperm’s viability. The article describes in depth how this analysis and interpretation is carried out.

Overall, this study provides an essential guide to studying sperm quality in large domestic animals, which is particularly significant in the animal reproduction industry, presenting techniques that offer in-depth analysis of sperm viability.

Cite This Article

APA
Hernández-Avilés C. (2025). Analysis of Motion Characteristics and Plasma Membrane Intactness (Viability) in Sperm from Domestic Animals. Methods Mol Biol, 2954, 241-259. https://doi.org/10.1007/978-1-0716-4698-4_14

Publication

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
NlmUniqueID: 9214969
Country: United States
Language: English
Volume: 2954
Pages: 241-259

Researcher Affiliations

Hernández-Avilés, Camilo
  • Equine Fertility Laboratory, Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA. chernandez@cvm.tamu.edu.

MeSH Terms

  • Animals
  • Male
  • Cell Membrane / metabolism
  • Cell Membrane / physiology
  • Spermatozoa / cytology
  • Spermatozoa / physiology
  • Flow Cytometry / methods
  • Sperm Motility / physiology
  • Semen Analysis / methods
  • Animals, Domestic
  • Horses
  • Cell Survival
  • Cryopreservation
  • Swine
  • Cattle

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