Detection of acrosome in mouse sperm incubated in vitro using fluorescence staining technique
The protocol for detecting acrosome in mouse sperm was developed by using two types of fluorescent dyes namely fluorescein conjugated Pisum sativum agglutinin (FITC-PSA) and rhodamine conjugated Lens culinaris agglutinin (TRITC-LCA). The results showed that mean percentages of acrosome-intact sper...
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| Main Authors: | , , |
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| Format: | Article |
| Published: |
2006
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| Subjects: | |
| Online Access: | http://eprints.um.edu.my/7857/ http://pkukmweb.ukm.my/msab/VOL%2035%20%20NO%201%20%20June%202006%20%20PDF/06%20%20Nooraain%20Hashim.pdf |
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| Summary: | The protocol for detecting acrosome in mouse sperm was developed by using two types of fluorescent dyes namely
fluorescein conjugated Pisum sativum agglutinin (FITC-PSA) and rhodamine conjugated Lens culinaris agglutinin
(TRITC-LCA). The results showed that mean percentages of acrosome-intact sperm incubated in vitro were
considerably low. After one-hour incubation there were 43.20±2.83, 31.81±8.44 and 30.90±4.15% for ICR, C57/6J
and F1 sperm, respectively. Intact acrosome in ICR sperm was significantly reduced (P<0.05) to 16.95±4.23% after
two-hour incubation, however, insignificant reduction (P>0.05) was found for C57/6J and F1
sperm, which were
30.03±2.06 and 20.93±3.81%, respectively. The results showed that both dyes FITC-PSA and TRITC-LCA suitably
stained the mouse sperm and produced insignificant difference (P>0.05) in the percentages of acrosome-intact sperm,
which were 40.03±4.20 and 49.79±4.63%, respectively. One-hour incubation was adequate to capacitate sperm in
vitro. Low acrosome intact due to extended period of incubation and morphologically abnormal sperm could
compromise in vitro fertilization rates. |
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