"Should we be using new tool of Flageliar Analysis and Sperm Tracking ( FAST)?"
INTRODUCTION: Can flagellar analyses be scaled up to provide automated tracking of motile sperm and does knowledge of the flagellar waveform provide new insight not provided by routine head tracking? The clinical gold standard for sperm motility analysis comprises a manual analysis by a trained professional, with existing automated sperm diagnostics
computer-aided sperm analysis (CASA)] relying on tracking the sperm head and extrapolating measures. It is not currently possible with either of these approaches to track the sperm flagellar waveform for large numbers of cells in order to unlock the potential wealth of information enclosed within.
SUMMARY: Using the software tool IFlagellar Analysis and Sperm Tracking (FAST)! Gallagher et al' analysed 176 experimental microscopy videos and have tracked the head and flagellum of 205 progressive cells in diluted semen (DSM), 119 progressive cells in a high-viscosity medium (HVM) and 42 stuck cells in a low-viscosity medium. The software tool in this manuscript has been developed to enable high-throughput, repeatable, accurate and verifiable analysis of the sperm &Filar beat. Unsc reened donors were recruited after giving informed consent. They described fully automated tracking and analysis of flagellar movement for large cell numbers. The analysis is demonstrated on freely motile cells in low- and high•-viscosity fluids and validated on published data of tethered cells undergoing pharmacological hyperactivation. Direct analysis of the flagella,- beat reveals that the CASA measure 'beat cross frequency' does not measure beat frequency; attempting to fit a straight line between the two measures gives R2R2 values of 0.042 and 0_00054 for cells in DSM and I-IVM, respectively. A new measurement, track centroid speed, is validated as an accurate differentiator or progressive motility. Coupled with fluid mechanics codes, waveform data enable extraction of experimentally intractable quantities such as energy dissipation, disturbance of the surrounding medium and viscous stresses. They provide a powerful and accessible research tool, enabling connection of the mechanical activity of the sperm to its motility and effect on its environment. The FAST software package has only been tested for use with negative phase contrast microscopy. Other imaging modalities, with bright cells on a dark background, have not been tested but may work. FAST is not designed to analyse raw semen; it is specifically for precise analysis of flagellar kinematics, as that is the promising area for computer use. flagellar capture will always require that cells are at a dilution where their paths do not frequently cross.
CONCLUSION: High-throughput flagellar waveform tracking and analysis enable measurement of experimentally intractable quantities such as energy dissipation, disturbance of the surrounding medium and viscous stresses, which are not possible by tracking the sperm head alone. Combining tracked flagella with mathematical modelling has the potential to reveal new mechanistic insight. By providing the capability as a free-to-use software package, ability to accurately quantify the flagellar waveform in large populations of motile cells will enable an abundant array of diagnostic, toxicological and therapeutic possibilities, as well as creating new opportunities for assessing and treating male subfertility.
SOURCE: https://www.indianfertilitysociety.org/fertility-news-august-2019-volume-11/
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