Fluorescene imaging of mouse sperm / Noorain H.
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...
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| Format: | Thesis |
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2012
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| Online Access: | http://studentsrepo.um.edu.my/3481/1/Chapter_6%2D2amac2010FINAL.pdf http://studentsrepo.um.edu.my/3481/ |
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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 C57BL/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 premature acrosome reaction during extended period of incubation may compromise in vitro fertilisation rates. |
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