Obstructed by hurdles in information extraction, handling and processing, computer-assisted sperm analysis systems have typically not considered in detail the complex flagellar waveforms of spermatozoa, despite their defining role in cell motility. Recent developments in imaging techniques and data processing have produced significantly improved methods of waveform digitization. Here, we use these improvements to demonstrate that near-complete flagellar capture is realizable on the scale of hundreds of cells, and, further, that meaningful statistical comparisons of flagellar waveforms may be readily performed with widely available tools. Representing the advent of high-fidelity computer-assisted beat-pattern analysis, we show how such a statistical approach can distinguish between samples using complex flagellar beating patterns rather than crude summary statistics. Dimensionality-reduction techniques applied to entire samples also reveal qualitatively distinct components of the beat, and a novel data-driven methodology for the generation of representative synthetic waveform data is proposed.

Collective swimming is evident in the sperm of several mammalian species. In bull (Bos taurus) sperm, high viscoelasticity of the surrounding fluid induces the sperm to form dynamic clusters. Sperm within the clusters swim closely together and align in the same direction, yet the clusters are dynamic because individual sperm swim into and out of them over time. As the fluid in part of the mammalian female reproductive tract contains mucus and, consequently, is highly viscoelastic, this mechanistic clustering likely happens in vivo. Nevertheless, it has been unclear whether clustering could provide any biological benefit. Here, using a microfluidic in vitro model with viscoelastic fluid, we found that the collective swimming of bull sperm in dynamic clusters provides specific biological benefits. In static viscoelastic fluid, clustering allowed sperm to swim in a more progressive manner. When the fluid was made to flow in the range of 2.43–4.05 1/sec shear rate, clustering enhanced the ability of sperm to swim upstream. We also found that the swimming characteristics of sperm in our viscoelastic fluid could not be fully explained by the hydrodynamic model that has been developed for sperm swimming in a low-viscosity, Newtonian fluid. Overall, we found that clustered sperm swam more oriented with each other in the absence of flow, were able to swim upstream under intermediate flows, and better withstood a strong flow than individual sperm. Our results indicate that the clustering of sperm can be beneficial to sperm migrating against an opposing flow of viscoelastic fluid within the female reproductive tract.

Plagued by hurdles in information extraction, handling, and processing, computer-assisted sperm analysis (CASA) systems have typically neglected the complex flagellar waveforms of spermatozoa, despite their defining role in cell motility. Recent developments in imaging techniques and data processing have produced significantly-improved methods of waveform digitisation. Here, we utilise these improvements to demonstrate that near-complete flagellar capture is realisable on the scale of hundreds of cells, and, further, that meaningful statistical comparisons of flagellar waveforms may be readily performed with widely-available tools. Representing the advent of high-fidelity computer-assisted beat-pattern analysis (CABA), we show how such a statistical approach can distinguish between samples using complex flagellar beating patterns rather than crude summary statistics. Dimensionality-reduction techniques applied to entire samples also reveal qualitatively-distinct components of the beat, and a novel data-driven methodology for the generation of representative synthetic waveform data is proposed.

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