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UTILITY OF LIGHT SCATTER IN THE MORPHOLOGICAL ANALYSIS OF SPERM
Citation:
Zucker, R., S. Perreault, AND K. Elstein. UTILITY OF LIGHT SCATTER IN THE MORPHOLOGICAL ANALYSIS OF SPERM. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-92/061 (NTIS PB92150754), 1992.
Description:
By using an Ortho flow cytometer to detect the forward light scatter from a red (helium-neon) laser, we were able to differentiate the morphologically diverse sperm nuclei of four animal species. ytograms composed of the axial light loss and forward red scatter signals revealed unique, but consistent sigmoid distributions that reflected not only interspecies morphological differences (shape, size), but variations in particle refractive index and orientation within the flow cell at the time of analysis. onsequently, we were able to use regional gating of the light scatter cytogram to minimize the influence of orientation on the resolution of fluorescence detection. In addition, we observed that sperm enlargement (resulting from chemically induced decondensation) coincided with a biphasic shift in light scatter that terminated, at a species-dependent rate, in attenuation of all measured forward scatter signals. hese results suggest that, without any special flow cell adaptations, flow cytometric light scatter parameters can be used to discriminate morphologically different sperm, to enhance the resolution of fluorescence measurements that may otherwise be confounded by variability in radial orientation, and to detect alterations In the rate of a biochemical/biophysical process (decondensation).
