Influence of sampling factors on canine sperm motility parameters measured by the Sperm Class Analyzer

Abstract

The aim of the present study was to investigate the effect of different technical settings and semen processing on sperm motility parameters measured by the Sperm Class Analyzer (SCA). Semen was collected from 3 dogs, pooled, and diluted in phosphate buffered saline and subsequently assessed by the SCA for the different sperm motility characteristics. The data were statistically analyzed by ANOVA and the repeatability was assessed by coefficient of variation (CV). After a principal component analysis, the reliability was determined with intraclass correlation coefficient (ICC). In experiment 1, the CV's were below 10% for all evaluated parameters. Significant differences (P < 0.05) were found between the different sperm concentrations (25, 50, and 75 x 10 6 spermatozoa/ml) in all of the motion parameters assessed, yielding the highest ICC (0.81) at 25 x 10 6 spermatozoa/ml. No significant differences (P > 0.05) in SCA read-outs were found between the number of microscopic fields captured (1, 2, 3, 4, or 5 fields), yielding the highest ICC (0.83) when 3 fields were captured. No significant differences (P > 0.05) in motility parameters were found between the number of cells analyzed in each field (20, 50, and 100 spermatozoa) with the exception of beat cross frequency. Reliability of the SCA was good (ICC = 0.71 to 0.90) for all motility measurements when 20 (ICC = 0.89) or 50 (ICC = 0.77) cells were captured in each field, but only just acceptable (ICC = 0.51 to 0.70) when 100 cells were counted (ICC = 0.67). The frame settings significantly (P < 0.05) influenced most of the measured motility characteristics. Scanning 60 frames at a frame rate of 30 Hz improved the reliability of the results (ICC = 0.92). In conclusion, we suggest that the measurements with the SCA are ideally performed at a sperm concentration of 25 x 10 6 spermatozoa/ml, counting at least 100 cells in three microscopic fields. We also propose that the SCA should analyze 60 frames at 30 Hz to yield consistent results of a set of measurements or a measuring instrument thus obtaining reliable motility results.

Ethanol-induced male infertility: Effects of aqueous leaf extract ofTetracarpidium conophorum

“…Sperm motility was evaluated by CASA using the Sperm Class Analyzer ® (SCA V.5.3, Microptic SL, Barcelona, Spain), as described by Dorado, Rijsselaere, Muoz‐Serrano, and Hidalgo (). The following sperm parameters were recorded: sperm concentration (total count; number of spermatozoa/rat × 10 6 ), total motility (%), progressive motility (%), nonprogressive motility (%), curvilinear velocity (VCL, μm/s), linear velocity (VSL, μm/s), average mean velocity (VAP, μm/s), linear coefficient (LIN, VSL/VCL × 100), straightness coefficient (STR, VSL/VAP × 100), wobble coefficient (WOB, VAP/VCL × 100), amplitude of lateral head displacement (ALH, μm) and beat cross‐frequency (BCF, Hz).…”

“…Sperm morphology was carried out as described by Dorado et al. (). Briefly, 10 μl of semen was dropped on a slide, and a thin feathered‐edge smear was made, and allowed to air‐dry fully for 5 min on a warm plate at 37°C.…”

Computer‐Assisted Sperm Analysis in Dogs and Cats: An Update after 20 Years

“…In dogs and cats, CASA was first described almost 20 years ago by Gu¨nzel-Apel et al (1993) and Stachecki et al (1993), respectively. Subsequently, numerous commercial CASA systems including the CellSoft computer videomicrography, Stro¨mberg-Mika Cell motion analyser, Hobson Sperm Tracker, Cell Track/s System, Hamilton-Thorne, Sperm Vision, Sperm Class Analyser were validated mainly in dogs and to a lesser extent in cats, both for research purposes and various clinical applications (Smith and England 2001;Verstegen et al 2002;Rijsselaere et al 2003;Scha¨fer-Somi and Aurich 2007;Dorado et al 2011).…”

Commensal Lactobacilli Enhance Sperm Qualitative Parameters in Dogs

“…Sperm kinematic parameters were evaluated using a computer-assisted sperm analysis system (Sperm Class Analyzer ® System version 6.4.0.93, Microptic, Barcelona, Spain), which included a Nikon Eclipse ci-L microscope (Nikon, Tokyo, Japan) with a 10 × phase-contrast objective, a heating stage at 37°C, with a frame rate set at 25 frames/s. Settings were adjusted according to the manufacturers recommendations ( 32 ), and a minimum of 500 cells in 5 random fields per sample were analyzed. For analysis, a sperm drop of 3 μl per sample was placed on Leja 20 μm chamber slides (Leja, Gynotec Malden, Nieuw Vennep, Netherlands) at 37° ( 33 ).…”