Background. Eye drops are most frequently used medications in ophthalmology. The carriage of pathogenic organisms to eyes through the agency of eye drops has presented a serious problem for several decades. The objective of this study was to determine the magnitude of contamination and pattern of antimicrobial resistance of in-use ophthalmic solutions. Method. A cross-sectional study was conducted at the Department of Ophthalmology, Jimma University Specialized Hospital (JUSH), Southwest Ethiopia, from June to December 2015. Samples from all ophthalmic solutions from outpatient department, operation theaters, and wards after an average duration of use of two weeks were taken. Samples were cultured and organisms were identified; antimicrobial susceptibility testing was performed using standard microbial identification techniques. The data were analyzed using SPSS software. Chi-square test was done and associations were taken as significant if P