Purpose: To test whether microwave oven irradiation can disinfect gypsum casts in compliance with current disinfection requirements, and to determine whether this procedure would be as effective as a validated method of chemical disinfection of impressions. Materials and Methods: In 2 in vitro experiments, samples of 5 irreversible hydrocolloid impressions of a disinfected acrylic resin model were contaminated with suspensions of recommended test organisms Staphylococcus aureus and Pseudomonas aeruginosa, respectively. In 1 in vivo experiment, 10 impressions were made of 10 subjects. All impressions were poured and the resulting casts cut in 2 halves. One half of each cast was exposed to 5 minutes of irradiation in a microwave oven at 2,450 MHz and 900 W. The other halves were left untreated as controls. In a second in vivo experiment, 10 impressions were disinfected by immersion in a 0.07% solution of NaOCl at pH of about 10 for 3 minutes, and then poured. All casts were incubated aerobically in Bacto tryptic soy broth at 37°C for 6 hours and assessed for bacterial growth by counting colony-forming units per milliliter (cfu/mL) of the culture. The results were analyzed using the Wilcoxon signed-rank test. Results: Untreated gypsum casts showed cfu/mL counts with a median log value of 6, while microwave-irradiated ones had median cfu/mL counts of 0. Casts poured from chemically disinfected impressions demonstrated cfu/mL counts with a median log value of 4. Conclusion: Under the described conditions, microwave-irradiated gypsum casts satisfy current disinfection requirements, but gypsum casts poured from chemically disinfected impressions do not.