Nooraain, H. (2012) Fluorescene imaging of mouse sperm / Noorain H. PhD thesis, University of Malaya.
Abstract
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.
Actions (For repository staff only : Login required)