Factors affecting cytoplasmic and nuclear structures of mouse gametes and embryos using immunocytochemistry technique / Nooraain Hashim

Experiment 1 was conducted with the main objective to establish ICC technique for morphological analysis of the mouse oocytes and embryos in local laboratory. Several tests were conducted to achieve specific aims which included; a) to study efficacy of the available instruments and system for detecting fluorescence signals from mouse oocytes, b) to compare efficiency of improved methods to reveal oocyte morphology. Fluorescence images of the meiotic spindles, cortical granules and nuclear materials were compared between sample preparations methods. The results showed that the methods employed were reliable to be used for detecting the meiotic spindle, cortical granules and nuclear materials. The microdroplet method used in staining procedure produced high number of total oocytes recovered as compared with poly-l-lysine coated cover slip method. Meanwhile, the appearance of meiotic spindle in oocytes was significantly affected by the method used during samples preparation. A significantly higher percentage of clear meiotic spindles images were obtained from zona intact and methanol fixed oocytes, which were 62.8% and 64.7%, respectively (p