The ability of fungi to adhere to host epithelial cells is a crucial step in the initiation of infection and pathogenesis of various fungal diseases (Martin et al, Future Med Chem 13:313–334, 2021). To study this adhesion process, a detailed model of fungal epithelial cell adhesion mimics the interaction between fungal cells and host tissue. The adhesion of pathogens is usually facilitated by microbial proteins, so-called adhesins, which bind to cell surface biomolecules such as protein receptors and glycans. In particular, the oral cavity provides surfaces to which fungal cells can adhere, such as buccal epithelial cells (BECs) (Martin et al, Eur J Med Chem 160:82–93, 2018). This model is based on the principle that fungal cell surface components, such as glycoproteins and adhesins, interact with specific receptors on the epithelial cell surface to enable adhesion. The extent of adhesion provides valuable information about the virulence of the fungal strain, the ability of the fungus to colonize host tissue and its potential to cause infection (Yang et al, Microbiol Res 169(11):803–810, 2014). This assay provides a simple but effective method to study host-pathogen interactions and can be used to test the efficacy of potential antifungal agents that interfere with fungal adhesion (Moyes et al, Virulence 6(4):338–346. 2015). Understanding these adhesion mechanisms is crucial for the development of strategies for the prevention and treatment of fungal infections. In this chapter, we present a detailed protocol for assessing fungal adhesion in which fungal cells are co-incubated with epithelial cells to allow cell-surface interactions. After a specific incubation period, the non-adherent fungal cells are washed away, and the adherent cells are visualized and quantified using staining techniques, such as crystal violet, which provide clear and reproducible results. These visual assays provide a straightforward method for assessing the extent of fungal adhesion to host cells in vitro.