Aim: The aim of this study was to detailed analysis of the donor corneal tissue using Eye Bank Specular Microscope (EBSM). Materials and Methods: Retrospective analysis was done for 408 donor corneas procured and processed between April 2016 and March 2020, at an Eye Bank in a Tertiary Care Hospital for Endothelial cell density (ECD), coefficient of variance (CV), and percentage of hexagonality (6A) and were compared across different age groups & gender of the donor tissue. Results: The mean age of the donors was 66.78 ± 17.27 (range 12–95 years). The mean ECD of the donor cornea was 1400.65 ± 627.01 cells/mm2; the average CV was 37.5 ± 18.2; and the average percentages of hexagonal cells were 44.9%. The cell count was significantly greater in the donor corneas in age groups of <20, 21-30 & 41-50 years when compared to rest of the age groups. There were no significant differences between the age groups on basis of CV and 6A. The ECD of the male donors (65%) was more than that of female donors (35%). Conclusion: ECD is significantly greater in younger age group as compared to older age groups; however, there were no significant differences in CV and 6A. Endothelial cell analysis by Eye Bank Specular Microscopy is a great tool for the grading of donor tissues, its subsequent utilization for corneal transplantation & prognostic value of the graft survival.