Abstract Background: Malaria is one of the killer diseases those caused by Plasmodium. Despite this, diagnosis is mainly based on clinical presentation having low specificity. Hence, prompt and reliable laboratory diagnosis is an important component of effective malaria case management and control. However, there is scarcity of qualified laboratory professionals for correctly diagnosing malaria using microscopy. Objective: The aim of this study was to evaluate the performance of laboratory professionals on malaria microscopy. Methods: A cross sectional study was conducted from January to March 2017 in Bahir Dar City. A purposive sampling technique was used to select health institutions. Then, all 87 laboratory professionals who were working at the selected health institutions were included in the study with a response rate of 95%. Data were collected and entered into SPSS version 23 for analysis. Assessment of performance of laboratory professionals on malaria microscopy was realized by computing the specificity, sensitivity, negative and positive predictive values. Percent of agreement was determined by kappa score. Bivariate logistic regression analysis was done to see the association between independent and dependant variable and p-value less than 0.05 at 95% CI was used to declare statistical significance. Results: Of the 87(95%) laboratory professionals participated in the study, 77(88.5%) had greater than two years experience and 49 (56.3%) of them took training once on malaria microscopy. From the total 696 positively and negatively validated panel slides, 616 (88.5%) were correctly detected by laboratory professionals. Similarly, of the 362 (52%) positively detected slides, 245(67.7%) were correctly identified. Laboratory professionals who were working at health centers had the highest performance on identification of Plasmodium species. The overall agreement of laboratory professionals’ performance on detection of malaria parasites and species identification were 88.5% and 72%, respectively. Conclusions: The present study showed that performance on identification of Plasmodium species was poor and none of them reported parasite density as per the standard. Therefore, regular training on species identification and strict follow up should be given by responsible bodies. Key words: Performance, Laboratory professionals, Malaria microscopy, Bahir Dar, Ethiopia.