The increasing number of endoscopic procedures performed and their increasing invasiveness mean that endoscopy of the gastrointestinal tract is associated with the risk of transmitting pathogenic microorganisms through infected equipment or contact with other patients and medical staff. In order to ensure protection of the health of both patients and medical staff, endoscopy laboratories should meet high hygiene standards. The results of tests of the microbiological cleanliness of surfaces and equipment of an endoscopic examination laboratory performed in the period from January to December 2019 at the Provincial Clinical Hospital No. 2 in Rzeszow were assessed retrospectively. Samples for testing were collected by swabbing from places where microbiological contamination was the most likely and cleaning was the most difficult. In the analyzed period, a total of 86 samples were collected for microbiological tests, of which positive results accounted for 6.9%. Positive results were obtained mainly from swabs collected from wet surfaces (66.7%). Most of the isolated microorganisms were Gram-negative bacteria (66.7% of all positive tests) and they were: Acinetobacter junii, Ralstonia pickettii, and Achromobacter denitrificans. The condition of the microbiological cleanliness of the surfaces and equipment of the endoscopic examination laboratory was satisfactory. A very low level of microbiological contamination of the tested items indicates occasional shortcomings in the decontamination processes. Since microorganisms isolated from the collected samples may be the cause of infection in patients and medical personnel, it is necessary to verify the decontamination procedures applied and to continue periodic microbiological monitoring of their effectiveness.