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Home / Equine Research Bank / Animals (Basel) / Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum F...
Animals : an open access journal from MDPI2020; 10(11); 1993; doi: 10.3390/ani10111993

Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields.

Abstract: In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISASD4C depths 10 and 20 µm, ISASD4C Leja 20 and drop displacement chamber (DDC), Spermtrack (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity-VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavior was observed along the CLC. However, all the kinematic variables had a higher value in a highly concentrated sample, in both chamber types. In conclusion, analyzing a minimum of nine fields at 250 fps from the centre to the edges in Spk10 chamber using a dilution of 30 × 10 sperm/mL offers the best choice for donkey computerised sperm motility analysis.
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Publication Date: 2020-10-29 PubMed ID: 33138237PubMed Central: PMC7692530DOI: 10.3390/ani10111993Google 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.

The research paper discusses optimizations for conducting donkey sperm motility analysis using the Computer Assisted Sperm Analysis (CASA)-Mot system, comparing different counting chambers and fields, and finding the best options for kinematic variables and frame rates.

Methodology

  • The researchers used twelve different ejaculate samples collected from six different jackasses.
  • They tested a variety of counting chambers for the CASA-Mot system, including Capillary loaded chamber (CLC), ISASD4C depths of 10 and 20 µm, Leja 20, and a drop displacement chamber (DDC) with Spermtrack (Spk) depths of 10 and 20 µm.
  • These various samples were then examined through a high-resolution camera which had a maximum capture capability of 500 frames per second (fps).

Findings

  • It was observed that the optimum frame rate (OFR), defined in relation to the curvilinear velocity (VCL), varied depending on the type of chamber being used.
  • The highest OFR achieved was 278.46 fps in the case of the Sperktrack depth 20 µm (Spk20) chamber.
  • Values for curvilinear velocity (VCL), straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH), and beat cross frequency (BCF) were higher in the drop displacement chamber (DDC) and chambers with depth 10 µm.
  • Within both the DDC 10 µm and 20 µm chambers, the sperm velocities (VCL, VSL, VAP), along with the lateral head displacement amplitude (ALH), decreased significantly from the centre to the edges. Meanwhile, wobble and BCF increased.
  • No defined behavior was observed along the CLC, although kinetic variable values were higher in samples with higher sperm concentration, regardless of chamber type.

Conclusion

  • After conducting these examinations, the researchers concluded that using a Spk10 chamber, dilution of 30 × 10^6 sperm/mL with a minimum of nine fields at 250 fps from the centre to the edges offers the best setup for conducting computer assisted sperm motility analyses on donkey sperm.

Cite This Article

APA
Gacem S, Catalán J, Valverde A, Soler C, Miró J. (2020). Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields. Animals (Basel), 10(11), 1993. https://doi.org/10.3390/ani10111993

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 10
Issue: 11
PII: 1993

Researcher Affiliations

Gacem, Sabrina
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, 08193 Bellaterra, Spain.
Catalán, Jaime
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, 08193 Bellaterra, Spain.
Valverde, Anthony
  • Costa Rica Institute of Technology, School of Agronomy, San Carlos Campus, 223-21001 Alajuela, Costa Rica.
Soler, Carles
  • Departamento de Biología Celular, Biología Funcional y Antropología Física, Universitat de València, 46100 Burjassot, Valencia, Spain.
Miró, Jordi
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, 08193 Bellaterra, Spain.

Conflict of Interest Statement

The authors declare no conflict of interest. Some of the authors (C.S. and D.B.) are employees for the company that manufacture the CASA-Mot system and the counting chambers used or collaborate through a collaborative agreement between the València University and the company that manufacture some of the components used here. In any case, it does not imply any conflict of interest, because no publicity (direct or indirect) is done in the manuscript. These components were just used in the same way we could use from other brands and the elements studied are not related with the brand but with physic and biological topics.

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Citations

This article has been cited 2 times.
  1. Yánez-Ortiz I, Catalán J, Delgado-Bermúdez A, Carluccio A, Miró J, Yeste M. Addition of Reduced Glutathione (GSH) to Freezing Medium Reduces Intracellular ROS Levels in Donkey Sperm.. Vet Sci 2021 Dec 2;8(12).
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  2. Gacem S, Valverde A, Catalán J, Yánez Ortiz I, Soler C, Miró J. A New Approach of Sperm Motility Subpopulation Structure in Donkey and Horse.. Front Vet Sci 2021;8:651477.
    doi: 10.3389/fvets.2021.651477pubmed: 34113670google scholar: lookup
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