In the past decade, the use of microtiter plates for microbial growth has become widespread, particularly in industry. In parallel, research in academia has provided a thorough insight into the complex relation between well dimensions, culture volumes, orbital shaking conditions and surface tension on the one hand, and oxygen-transfer rates and degrees of mixing on the other. In this review, I will discuss these fundamental issues and describe the current applications of microtiter plates in microbiology. Microtiter plates can now be considered a mature alternative to Erlenmeyer shake flasks.