Adhesion to host tissue represents one of the first steps during the early phase of fungal infections. In order to mediate pathogenesis in the infected host, this process is crucial for colonization and subsequent penetration of the respective tissue. In vivo analyses of the adhesion process in whole organisms are limited because of difficulties in providing reproducible and comparable conditions in the host environment. Therefore, in vitro assays provide the opportunity to study such processes under more defined conditions thus allowing for the analysis of events that are involved in more detail. Here we describe an in vitro adhesion assay making use of human epithelial cell lines to study fungal associations with host epithelia. This assay not only is suited to determine the rate of adhesion in a time-dependent manner but also facilitates global transcriptional profiling in order to determine the fungal response during adhesion at the molecular level.