Indentification of Caspase 3 Intensity in Kacang Goat Oocyte Post Vitrification using Confocal Laser Scanning Microscopy

The research was aimed to find out caspase 3 intensity in Kacang goat oocyte post vitrification. The high caspase intensity will affect the viability of post thawing oocytes so that it will affect the oocyte quality and fertilization rate. The intensity of caspase 3 expression occurs because of the bond between caspase 3 with the Flou 3 probe so it will emit a fluorescent color, which is captured by the CLSM microscope device. The benefits of this research were to measure the intensity of caspase 3 quantitatively so as to determine the oocyte quality post thawing and determine the quality of frozen oocytes as a source of gametes for fertilization purposes. There were 2 groups in the research; Kacang goat fresh oocyte and kacang goat oocyte that was vitrified (each group consisted of 5 replications). The procedure of this study including oocyte collection, oocyte vitrification, frozen oocyte warming and Caspase 3 intensity examination were labeled with anti rabbit Fluorescein Isothiocyanate (FITC) secondary antibodies and measurement with Confocal Laser Scanning Microscopy (CLSM). Caspase 3 intensity of both groups was observed using CLSM. The results showed that caspase 3 intensity in the oocyte group which had been frozen by vitrification was higher significant than fresh oocytes (P < 0.05). The research concluded that caspase 3 intensity of frozen oocyte post vitrification was higher than that of fresh oocyte.