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Biology of reproduction1986; 34(1); 127-138; doi: 10.1095/biolreprod34.1.127

Assessment of spermatozoal function using dual fluorescent staining and flow cytometric analyses.

Abstract: Spermatozoa from bulls, boars, dogs, horses, mice, and men were examined using a fluorogenic stain consisting of the membrane-permeant substrate carboxyfluorescin diacetate (CFDA) and the relatively membrane-impermeant nuclear stain propidium iodide (PI). Three distinct populations of spermatozoa were discernible in samples from each species upon microscopic examination. Individual spermatozoa, presumed to be viable because of their motility, retained products of the fluorescein chromophore throughout the cell. A second population of spermatozoa in which the nuclei stained red with PI retained the green fluorescein fluorophore mainly in the acrosome. A third population, presumed to be degenerate spermatozoa, possessed only red fluorescent nuclei. These populations were quantified using dual parameter flow cytometry in 14 samples of cryopreserved bovine spermatozoa for which fertility and seminal quality data were available. Flow cytometric analyses were highly correlated with other seminal quality measurements. Sequential flow cytometric analyses provided the ability to rapidly quantitate changes in specific fluorescently stained populations. The ability to make rapid quantitative measurements should allow development of new and presumably more reliable information on the functional aspects of spermatozoa.
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Publication Date: 1986-02-01 PubMed ID: 3955132DOI: 10.1095/biolreprod34.1.127Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

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.

The research explored the use of dual fluorescent staining and flow cytometric analyses to assess the functionality of spermatozoa from various species. The study demonstrated a strong correlation between the results from this technique and other seminal quality measurements, suggesting a possible new approach for quick and reliable sperm quality assessment.

Methodology

  • The sperm samples used in this study were sourced from several species including bulls, boars, dogs, horses, mice, and humans.
  • A dual fluorescent stain was used for the examination of spermatozoa. This stain comprised of carboxyfluorescein diacetate (CFDA), a substrate that can diffuse through cell membranes, and propidium iodide (PI), a nuclear stain unable to easily permeate membranes.
  • Three unique populations of spermatozoa became apparent upon microscopic assessment based on fluorescent staining patterns.
  • The research also performed flow cytometric analyses on 14 samples of cryopreserved bovine sperm, which were correlated with fertility and seminal quality data for the same samples.

Findings

  • The study identified three distinct populations of spermatozoa. The first group were thought to be viable sperm due to its motility and retention of products from the fluorescein chromophore (CFDA) throughout the cell.
  • A second population of sperm was identified, their nuclei were stained red by the PI, and were noted to retain the green fluorescein fluorophore mostly in the acrosome, an important part of the sperm cell that contains enzymes required for penetrating the egg during fertilization.
  • The third population presumably consisted of degenerate spermatozoa that only exhibited red fluorescent nuclei, staining positively with PI, indicating compromised membrane integrity.
  • The study found a high correlation between the flow cytometric analyses results and other seminal quality measurements, suggesting that the flow cytometric analyses could quickly provide reliable quantitative information on sperm functionality.

Implications

  • This research suggests that the dual fluorescent staining protocol and flow cytometric analyses may be a reliable, quick method for assessing the functional aspects of spermatozoa, which could potentially be advantageous in various fields such as fertility clinics and animal breeding.

Cite This Article

APA
Garner DL, Pinkel D, Johnson LA, Pace MM. (1986). Assessment of spermatozoal function using dual fluorescent staining and flow cytometric analyses. Biol Reprod, 34(1), 127-138. https://doi.org/10.1095/biolreprod34.1.127

Publication

Biology of reproduction
ISSN: 0006-3363
NlmUniqueID: 0207224
Country: United States
Language: English
Volume: 34
Issue: 1
Pages: 127-138

Researcher Affiliations

Garner, D L
    Pinkel, D
      Johnson, L A
        Pace, M M

          MeSH Terms

          • Acrosome / physiology
          • Animals
          • Cattle
          • Dogs
          • Flow Cytometry / instrumentation
          • Flow Cytometry / methods
          • Fluoresceins
          • Horses
          • Humans
          • Male
          • Species Specificity
          • Sperm Count
          • Spermatozoa / cytology
          • Spermatozoa / physiology
          • Swine

          Citations

          This article has been cited 18 times.
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