The in vitro antimicrobial and cytotoxic concentrations of glutaraldehyde and formocresol were determined. Minimal antimicrobial concentrations of these two agents against selected microbial flora reported in carious primary teeth were 3.125% for glutaraldehyde and 0.75% for formocresol. At a doubling of these concentrations, most organisms, except Candida albicans, Staphylococcus epidermidis, and Streptococcus mutans, were killed by both substances in 30 seconds. Cytotoxicity was evaluated on tissue cultures of pulp fibroblasts and HeLa cells at minimal cidal concentrations and at 10- and 100-fold dilutions. Exposure of pulp fibroblasts and HeLa cells to formocresol and indirect exposure to vapors caused the cells to become atrophic and to form a less dense tissue pattern. Cells directly exposed to glutaraldehyde retained their normal cell shape and tissue pattern, whereas cells indirectly exposed to vapors continued to proliferate. These data showed effective antimicrobial activity at concentrations of 3.125% glutaraldehyde and 0.75% formocresol and suggested that glutaraldehyde may exert a less cytotoxic effect on the immediate and surrounding tissues when used as a pulpotomy agent.