Owing to its ability to form biofilms on implanted medical devices, the fungal pathogen Candida albicans causes frequent infections in humans. A hallmark of C. albicans biofilms is the presence of two types of cells, budding yeast cells and growing hyphae, which are bound together and embedded in extracellular matrix material. Although cell-cell adhesion is critical to biofilm formation, architecture, and cohesion, we know little about the fundamental forces behind this interaction. Here, we use single-cell force spectroscopy to quantify the forces engaged in yeast-hyphae adhesion, focusing on the role of Als (agglutinin-like sequence) proteins as prototypes of cell adhesion molecules. We show that adhesion between individual yeast and hyphal cells involves strong, short-range cohesive interactions (1.1 +/- 0.2 nN; 86 +/- 33 nm) and weak, long-range tether interactions (0.4 +/- 0.2 nN; 234 +/- 81 nm). Control experiments demonstrate that these interactions originate from cell surface proteins that are specific to C. albicans. Using mutant strains deficient for Als expression, we find that Als3 proteins, primarily expressed on the germ tube, play a key role in establishing strong cohesive adhesion. We suggest a model in which cohesive adhesion during biofilm formation originates from tight hydrophobic interactions between Als tandem repeat domains on adjacent cells. When subjected to force, the two interacting cell surfaces detach, but the cell bodies remain tethered through macromolecular extensions. Our results represent the first direct, noninvasive measurement of adhesion forces between interacting fungal cells and provide novel insights into the molecular origin of the cohesive strength of fungal biofilms.