X-Message-Number: 26246
From: "Basie" <>
Subject: Some damage not due to freezing and thawing process
Date: Wed, 25 May 2005 19:10:30 -0400

Theriogenology 54: 1075 - 1086
Significance of plasmalemma disruption in bovine and equine spermatozoa.
Abraham-Peskir, J. V., Chantler, E., and Uggerhoj, E.
Institute for Storage Ring Facilities, University of Aarhus, Denmark. 


We have investigated fresh and cryopreserved bovine and equine spermatozoa 
using light and transmission soft X-ray microscopy. Spermatozoa were 
examined, in the presence or absence of semen, after using Percoll gradient 
centrifugation and re-suspending in medium. X-ray microscopy provided high 
resolution (30 nm) transmission images of whole cells in solution with high 
contrast, while retaining the simple preparation techniques used in light 
microscopy. We demonstrated translucent, membrane-bound vesicles in the 
acrosomal and midpiece regions that were similar in size and we noted their 
incidence in both fresh and frozen-thawed material from both animals. The 
vesicles were formed by the separation and expansion of the plasmalemma away 
from the underlying structure but were not caused by the freeze-thaw 
process. We suggest that these structures form part of the normal 
ultrastructure of spermatozoa and are damaged during preparation of the 
samples for transmission electron microscopy, resulting in a structure 
previously and incorrectly identified as damaged by the freezing and thawing 
process

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