Background: Candida albicans is a low level commensal organism in normal populations with the continous potential to expand and cause a spectrum of clinical conditions. Methodology/Principal Findings: Using ex vivo human organ cultures and populations of primary human cells, we have development several related experimental systems to examine early-stage interactions between C. albicans and mucosal surface. Experimental have been conducted both with exogenously added C. albicans and with overtly normal human mucosal surfaces supporting pre-existing infections with natural isolates of Candida. Under different culture conditions, we have demonstrated the formation of C. albicans colonies on human target cells and filament formation, equivalent to tissue invasion. Conclusion/Significance: These organ culture system provide a valuable new resource to examine the molecular and cellular basis for Candida colonization of human mucosal surfaces.