[Observation of the acrosome reaction of human spermatozoa using the combined preparation with triple stain technique and scanning electron microscopy].
The combination of triple stain technique (TST) and scanning electron microscopy (SEM) was used to investigate the reliability of TST in observing the acrosome reaction. The TST preparation was followed by SEM to observe the same spermatozoa by both methods. We compared the results obtained with TST at various pH of rose Bengal solution to those with SEM. 1. In our method, the acrosome reaction on the same spermatozoa was easily examined by both TST and SEM. 2. The fine surface structure of the sperm head was not affected by the TST procedure, because the percentages of acrosome non-reacted (ANR) and acrosome reacted (AR) sperm were similar when spermatozoa from the same semen were examined by SEM or SEM after TST (23.0%, 77.0% VS. 24.5%, 75.5%, respectively). 3. When results obtained with TST at various pH of rose Bengal solution were compared to those of SEM after TST, the accuracy was 70.5% at pH 4.3, 95.9% at pH 5.3 and 61.8% at pH 6.3. Sensitivity and specificity were 100%, 21.4% at pH 4.3, 94.5%, 97.6% at pH 5.3 and 38.7%, 97.5% at pH 6.3 respectively. From these results, it can be concluded that TST with pH 5.3 rose Bengal solution is a reliable and practical method for detecting the acrosome reaction of human spermatozoa.
Bandoh
R
R
Department of Obstetrics and Gynecology, School of Medicine, University of Tokushima.
Kagawa
T
T
Yamano
S
S
Aono
T
T
D000177:Acrosome / Q000648:ultrastructure*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D008855:Microscopy, Electron, Scanning; D012395:Rose Bengal; D013094:Spermatozoa / Q000648:ultrastructure*; D013194:Staining and Labeling
D012395:Rose Bengal