If not adequately cleaned, rigid nasal endoscopes (RNEs) have the potential to cause iatrogenic cross-contamination. To test the efficacy of various disinfection methods in reducing bacterial load on RNEs in vitro. In vitro model. Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, and Haemophilus influenzae contamination was separately induced on RNEs in vitro. Two experimental sets were completed. The RNEs were disinfected using the following protocols: 30-second scrub with antimicrobial soap (ABS) and water, 30-second scrub with 70% isopropyl alcohol (IA), 30-second scrub with ABS followed by 30-second scrub with IA, 30-second scrub with germicidal cloth, isolated 5-minute soak in an enzymatic soap solution, 5- and 10-minute soaks in ortho-phthalaldehyde, 0.55%, solution (Cidex OPA), and isolated 30-second rinse with tap water, all with 30-second precleaning and postcleaning rinses with tap water. Two sets of experiments (experiment sets A and B) were carried out with a 30-second tap water rinse after inoculation of each RNE. This was followed by immediate cleaning in set A and a 1-hour air-dry delay in set B. Otherwise there were no differences in the disinfection protocols between sets for each method noted. Effectiveness of various disinfection protocols in cleaning rigid nasal endoscopes experimentally inoculated with bacteria commonly found in the upper aerodigestive tract. Positive cultures following disinfection indicated ineffective or incomplete disinfection. Most cleaning methods were effective in eliminating S aureus, S pneumoniae, and H influenzae from the scopes following experimental contamination. Continued growth of P aeruginosa was found after all of the disinfection trials in experiment set A with the exception of a 10-minute immersion in Cidex OPA, and in set B except for the 10-minute Cidex OPA immersion and ABS plus IA trials. Most cleaning methods used in our trials appear to properly disinfect RNEs after in vitro inoculation with S aureus, S pneumoniae, and H influenzae. However, it appears that disinfectants may be less effective in cleaning rigid scopes experimentally inoculated with P aeruginosa. There is a paucity of published data regarding cross-contamination during rigid nasal endoscopy, and these results should guide future studies and to some extent practice to avoid iatrogenic spread of contamination.