Live sperm
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Live sperm is assessed by employing the differential staining technique as described by Hancock (1951); 5 per cent eosin-Y and 10 per cent nigrosin solution in 2.9 per cent sodium citrate solution is utilized for staining.
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A drop of eosin, two drops of nigrosin and a small drop of semen are placed on a slide and mixed.
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This is taken on the edge of a slide and pulled across the top of another slide leaving a smear.
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Allow it to dry in air.
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200 spermatozoa are counted under oil immersion at a magnification of 1000X in different areas of smear and the live sperm percent is calculated.
Interpretation
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Unstained sperms are considered as LIVE.
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Sperms stained pink by eosin are considered as DEAD.
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Partially stained sperms are considered as DEAD.
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- Several procedures have been developed for objective (unbiased) evaluation of sperm motility: time-lapse photomicrography, frame-by-frame playback videomicrography, spectrophotometry, and computerized analysis.
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Computer assisted semen analysis (CASA) system are used in reference laboratories as an objective means of assessing sperm motility.
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Sperm descriptions with CASA includes: curvilinear velocity, average path velocity, straight line velocity, amplitude of lateral head displacement, linearity, beat cross frequency, mean angular displacement, area or tracked sperm head and time each spermatozoa is tracked.
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Flow cytometry can detect viable sperm based on binding of fluorescent dyes (SYBR-14 and Propidium Iodide).
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Time lapse photomicroscopy permits visualization of sperm tracks.
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Last modified: Wednesday, 6 June 2012, 2:25 PM
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