BACKGROUND Microbial keratitis is a potentially vision threatening state requiring prompt diagnosis & treatment, and is a leading cause of preventable blindness. We wanted to study the spectrum and sensitivity of microbiological isolates from patients of microbial keratitis at a tertiary care centre of a metropolitan city. METHODS Descriptive data collection and analysis was carried out in a tertiary care centre over a period of 3 years. The cases were the grouped based upon their positivity in microscopy and culture results and correlated with clinical data. RESULTS 258 clinical cases were studied, of which 173 cases were bacterial keratitis, and 85 were fungal keratitis. Of 173 (67.05%) bacterial cases, 68 (39.30%) cases showed positive findings on gram stain, and 25 (36.76%) of these cases were confirmed by culture. Of 85 (32.94%) fungal cases, 37 (43.53%) cases were positive on KOH mount & 21 (56.76%) of these cases were confirmed by culture. CONCLUSIONS Incidence of bacterial keratitis is higher than fungal keratitis. Emphasis should be on using microbiological diagnosis as an important tool to aid the ophthalmologist in appropriate management of these cases.