OBJECTIVE To compare the effectiveness of manual and automated methods for cleaning laparoscopic instruments. DESIGN Experimental laboratory study. METHODS We evaluated 4 methods of cleaning laparoscopic instruments: (1) manual-only cleaning and rinsing with potable tap water; (2) manual cleaning and rinsing with potable tap water, followed by ultrasonic cleaning without rinsing; (3) manual cleaning and rinsing with potable tap water followed by ultrasonic cleaning and rinsing with potable tap water; and (4) manual cleaning and rinsing with potable tap water, followed by ultrasonic cleaning and rinsing: first with potable tap water and then with sterile distilled water. Organic residues of protein, hemoglobin, and carbohydrates were evaluated using spectrophotometry. RESULTS The various cleaning methods tested did not result in statistically significant differences (P>.05) in the levels of investigated organic residues. CONCLUSIONS All cleaning and rinsing methods tested were found to be effective in reducing the levels of organic residues on laparoscopic instruments. Thus, there is no advantage gained by supplementing manual-only cleaning with automated ultrasonic cleaning, nor was there a difference between rinsing with potable tap versus sterile distilled water. Infect Control Hosp Epidemiol 2018;39:58-63.