Real-time imaging of fungal infections is becoming integral to the study of host-pathogen interactions, as it allows monitoring of the spatial and temporal progression of pathogen growth or of the host response in a single animal as well as reducing the number of animals used to obtain significant data. We present different applications of a novel luciferase reporter gene constructed from the coding sequences of the Candida albicans PGA59 gene, encoding a GPI-linked cell wall protein, and the Gaussia princeps luciferase gene. Upon addition of the coelenterazine substrate, light produced by the surface-exposed luciferase can be used to quantify gene expression from a variety of C. albicans promoters as well as monitoring cutaneous, subcutaneous, and vaginal infections.