Summary
Methods are given for the preparation and staining of human spermatozoa for flow cytometric DNA measurements. Using agents for the reductive cleavage of disulfide crosslinks and suitable proteolytic enzymes and effective decondensation of the sperm chromatin and a DNA-proportional uptake of fluorochromes is achieved. Thus reliable and precise measurements of the relative DNA content of human spermatozoa are possible and the two subpopulations of haploid spermatozoa can be distinguished according to the difference in their DNA content.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Barlogie, B., Zante, J., Drewinko, B., Schumann, J., Büchner, T., Göhde, W., Hart, J.S., Johnston, D.A.: The use of mithramycin in pulse-cytophotometry. In: Pulse cytophotometry. W. Göhde, J. Schumann and Th. Büchner (eds.) Vol. II, pp. 125–136. Ghent: European Press Medikon 1976
Crissmann, H.A., Steinkamp, J.A.: Comparison of quantitative fluorescent staining methods for flow-system analysis of DNA, protein, and nuclear-to-cytoplasmic size in cultured cells, tumor cells, and human gynecological specimens. In: Pulse cytophotometry. W. Göhde, J. Schumann and Th. Büchner (eds.) Vol. II, pp. 112–124. Ghent: European Press Medikon 1976
Dean, P.N., Pinkel, D., Mendelsohn, M.L.: Hydrodynamic orientation of sperm heads for flow cytometry. Biophys. J. 23, 7–13 (1978)
Gall, W.E., Ohsumi, Y.: Decondensation of sperm nuclei in vitro. Exp. Cell Res. 102, 349–358 (1976)
Gledhill, B.L., Lake, S., Steinmetz, L.L., Gray, J.W., Crawford, J.R., Dean, P.N., Van Dilla, M.A.: Flow microfluorometric analysis of sperm DNA content: effect of cell shape on the fluorescence distribution. J. Cell. Physiol. 87, 367–376 (1976)
Meistrich, M.L., Göhde, W., White, R.A., Schumann, J.: Resolution of X and Y spermatids by pulse cytophotometry. Nature 274, 821–823 (1978)
Mendelsohn, M.L., Mayall, B.L., Bogart, E., Moore, D.H., Perry, B.H.: DNA contents and DNA-based centromeric index of the 24 human chromosomes. Science 179, 1126–1129 (1973)
Milette, C.F., Spear, P.G., Gall, W.E., Edelmann, G.M.: Chemical dissection of mammalian spermatozoa. J. Cell Biol. 58, 662–675 (1973)
Schweizer, D.: Reverse fluorescent chromosome banding with chromomycin and DAPI. Chromosoma 58, 307–324 (1976)
Schweizer, D., Ambros, P., Andrle, M.: Modification of DAPI banding on human chromosomes by prestaining with a DNA-binding oligopeptide antibiotic, distamycin A. Exp. Cell Res. 111, 327–332 (1978)
Sumner, A.T., Robinson, J.A., Evans, H.J.: Distinguishing between X, Y, and YY-bearing human spermatozoa by fluorescence and DNA content. Nature (New Biol.) 229, 231–233 (1971)
Van Dilla, M.A., Gledhill, B.L., Lake, S., Dean, P.N., Gray, J.W., Kachel, V., Barlogie, B., Göhde, W.: Measurement of mammalian sperm deoxyribonucleic acid by flow cytometry. Problems and approaches. J. Histochem. Cytochem. 25, 763–773 (1977)
Zante, J., Schumann, J., Barlogie, B., Göhde, W., Büchner, Th.: New preparating and staining procedures for spezific and rapid analysis of DNA distributions. In: Pulse cytophotometry. W. Göhde, J. Schumann and Th. Büchner (eds.). Vol. II, pp. 97–106. Ghent: European Press Medicon 1976
Zante, J., Schumann, J., Göhde, W., Hacker, U.: DNA-fluorometry of mammalian sperm. Histochemistry 54, 1–7 (1977)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Otto, F.J., Hacker, U., Zante, J. et al. Flow cytometry of human spermatozoa. Histochemistry 61, 249–254 (1979). https://doi.org/10.1007/BF00508445
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00508445