The use of CO2 under pressure (dense CO2) is one of the most promising techniques to achieve cold pasteurization and/or sterilization of liquid and solid materials, and is likely to replace or partially substitute currently and widely applied thermal processes. Although the ability of CO2 to inactivate microorganisms has been known since the 1950's, only within the last 15 years it has received special attention, and the scientific and economic interest towards practical applications is presently growing more and more. Here we collect and discuss the relevant current knowledge about the potentials of dense CO2 as a non-thermal technology in the field of microbial inactivation. We summarize the state of the art, including definitions, description of the equipment, relevant applications, in both simple suspensions and complex media, for the treatment of a wide range of microorganisms in both liquid and solid substrates. Finally, we also summarize and discuss the different hypotheses about the mechanisms of inactivation © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng84: 627–638, 2003.