Life optic slide

We used total and progressive motility post-thaw motility as the main metrics in models to analyze and interpret our multiple statistics. We used a Sperm Class Analyzer (Microptic, Barcelona, Spain) that captures video at 50 frames per second. The diluted semen (2.5 µL) was placed on a pre-warmed chamber slide (33 °C, Life optic slide, 20 µM depth chamber), and motility metrics were determined using a phase-contrast microscope (Nikon, Mississauga, ON, Canada) with a 10× objective at 33 °C. The following cut-offs were applied for all CASA analyses: the range for particle size was defined as 8 to 150 µm²; VCL was used to characterize immotile cells (VCL < 40 µm/sec), slow cells (VCL between 40 and 80 µm/s), medium cells (VCL between 80 and 150 µm/s) and fast cells (VCL > 150µm/s); and progressive motility was defined as >85% STR. A minimum of 300 spermatozoa from 8 fields in 50 frames of video were acquired for each analysis.

For the first 17 generations, we used post-thaw motility as the optimization metric for our model to get our model within a comparable range to our control extender medium post thaw recovery. At generation 18, we changed to relative total motility as our optimization metric to give more power to our post thaw metrics by controlling for individual bull variability. The relative total motility was calculated by taking a ratio of algorithmic total motility over control total motility of each sample. Three additional iterations were performed using the new input parameter.
Motility was evaluated using the Sperm Class Analyzer (Microptic, Spain) capturing videos at 50 frames per second during one second^{48 (link)}. The diluted semen (2.5 µl) was placed on a pre-warmed chamber slide (37 °C, Life optic slide, 20 µM depth chamber) and motility characteristics were determined using a phase-contrast microscope (Nikon, Canada) with a 10 × objective at 33 °C.