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Technical Report: Advancing Sperm Quality Assessments for Amphibians: Developing Flow Cytometry for Precise Evaluation of amphibian sperm viability

Leah Jacobs , Talisin Hammond, Natalie Calatayud, Patricia Byrne, Tom Jensen

Abstract

Aims: In the past decade flow cytometry has become a useful tool for evaluating cellular viability characteristics for non-domestic animals. This technology has the potential to vastly improve sperm quality assessments, concentration counts and cell sorting in a more time efficient and reliable manner. The study aimed to validate the efficacy of using flow cytometry for amphibian sperm by comparing its results with those obtained through traditional means of sperm quality assessment. Methods: Sperm samples were collected from testes macerates of the African clawed frog (Xenopus laevis) and subjected to both flow cytometry and microscopy analyses. Flow cytometry allowed for the simultaneous assessment of sperm viability and concentration using fluorescent probes, while microscopy provided a traditional means of assessing sperm characteristics. Key Results: Sperm concentrations measured by flow cytometry and fluorescent microscopy were highly correlated, though flow cytometry methods estimated significantly higher concentrations. Sperm viability as measured by flow cytometry and fluorescent microscopy methods were not significantly correlated but were also not significantly different. Conclusions: Although flow cytometry overestimated concentration and live/dead assessments, the discrepancies were slight enough to indicate that flow cytometry can still be a valuable method for assessing amphibian sperm. Implications: These results validate the utility of flow cytometry as a reliable tool for assessing amphibian sperm viability and concentration, offering a promising alternative to traditional, time-consuming methods.

RD24117  Accepted 16 December 2024

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