Biol Reprod Keystone Symposia Conference on Frontiers in Reproductive Biology & Regulation of Fertility.
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Biology of Reproduction, Vol 3, 8-12, Copyright © 1970 by Society for the Study of Reproduction

A Fluorescence Staining Method for Studying Sperm Membranes

R. J. ERICSSON 1, and D. A. BUTHALA 1

1 Research Laboratories, The Upjohn Company, Kalamazoo, Michigan 49001


Tetracycline HCl (T-HCl) is a supravital stain for mammalian sperm. As measured by fluorescence microscopy, T-HCl stains live and dead sperm but not with equal intensity. Dead sperm fluoresce more brightly than live sperm (except human) due to a greater uptake of T-HCl. The equatorial segment is visible on live sperm of rabbit, bull, and monkey but not human. This region is seen most easily on dead sperm, especially those which have lost their acrosomes. The plasma membrane can and does partially separate from heads of some live human sperm. Most, if not all, rabbit, bull, and monkey sperm have plasma-membrane separation from the head as a postmortem occurrence. This aging process starts with a ballooning of membranes around the anterior region of the head and culminates, in some sperm, with loss of the acrosome. Membrane separation can include both plasma and outer acrosomal membranes; the plasma membrane infrequently becomes separated from the entire head which reveals the separated acrosomal membrane. A distinct advantage of this technique is the study of live sperm and potential membrane differences in various conditions of fertility.

Submitted on September 3, 1969






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Copyright © 1970 by the Society for the Study of Reproduction.