Attempts to devise an antimicrobial approach to combating dentomicrobial infections such as periodontal diseases continue to be hampered by the lack of a relevant in vitro method for determining the susceptibility of suspected periodontopathogens to topically applied antimicrobial agents. Proposed here is a novel in vitro method called the membrane transfer technique, which acknowledges those aspects unique to localized pathogenic infections, particularly those associated with anaerobic bacteria. Bacterial lawns representing six suspected periodontopathic bacteria were prepared on membranes and then placed in contact wth different concentrations of antimicrobial agents for 5 min. After incubation for 12 to 24 h, MBCs were determined with the aid of a tetrazolium chloride indicator. Four antimicrobial agents (chlorhexidine, iodine, stannous fluoride, and sodium fluoride) were used to test the applicability of the proposed in vitro method. MBCs were derived for each agent except sodium fluoride against all or most of the six bacterial strains tested. The proposed method may also be useful for examining the bactericidal action of topically applied antimicrobial agents against nonoral infections.