External eye disease which result in corneal scarring are an important cause of blindness in Bangladesh and at the Chittagong Eye Infirmary and Training Complex (EITC) over 200 cases of suppurative keratitis are managed each year. We reviewed the records of 127 cases of microbial keratitis to determine the relative contributions of Gram stain and culture to diagnosis of the causative organism. There were 107 culture-proven cases of microbial keratitis amongst the 127 patients in this study. Gram stain was positive in 89 cases which represents 70% of the total and 83% of all culture-proven cases. Streptococcus pneumoniae and Pseudomonas sp were the commonest bacteria isolated and Aspergillus sp and Fusarium sp the commonest fungi. In 20 cases (16%) no organism was isolated on Gram stain or culture. Our results support the use of both Gram stain and culture in isolation of the causative organism in cases of suppurative keratitis in Bangladesh. However the low cost of Gram stain and its useful recovery rates for both bacteria and fungi support its use as an initial investigation for microbial keratitis at the secondary level of eye care in rural Bangladesh.