Cryopreservation of Zebrafish Ovarian Follicles Essay

Cryopreservation of Zebrafish Ovarian Follicles - Essay Example The technologies in cryopreservation have undergone tremendous advances over the last decade. In addition, cryopreservation of oocytes and embryos is also being used as an effective means of treating infertility. In this technology, the clinical application seeks to ensure the optimal survival of embryos and oocytes that are subsequently thawed and stored for transfer . The aim of this practical experimentation is to compare the slow-cooling procedures with vitrification to analyze and evaluate the most effective and safest procedure as well as to endorse suitable recommendation for the adoption of best practices. To do this, it is necessary to test and calculate the viability for control, slow-cooling and vitrification samples. Determining the number of cells in the culture is also important for standardising culture conditions and performing accurate quantitation experiments . The use of viability test with hemacytometer and typan blue staining will enable us to determine the cell number, the correctness of which is inevitable for accurate test results. Live cells appear colourless and bright under phase contrast, while the dead cells sustain blue stains and are non-refractive. To facilitate accuracy and consistency of cell counts, we have used a viability counting system. This involves counting viable, live and dead cells in one or more large corner squares and recording the cell counts. In order to obtain an accurate cell count, 40 to 70 cells will be counted during the test phase. Therefore, it may be necessary to count more than one large corner square. The controlled technique, which is the conservative method used for the purpose of cryopreservation of cells and tissues, is based on the slow-cooling approach. It needs to be appreciated that a large number of non-sensitive cells can be preserved in liquid nitrogen with little damage through slow-cooling