Visualizing Cell Wall Components in Candida albicans

Overnight cultures of C. albicans SC5314 were incubated in YPD medium with or without 11g at 30°C, favoring yeast-form growth. For hyphal growth, C. albicans SC5314 (1 × 10⁶ cells/mL) was incubated in Spider medium with or without 11g at 37°C for 3 h. Cells were then washed three times with PBS and then stained with Concanavalin A (ConA; Life Technologies, Carlsbad, CA, United States) to visualize mannan, anti-β-glucan primary antibody (Biosupplies, Bundoora, Australia) followed by Cy3-labeled secondary antibody (Life Technologies) to visualize β-(1, 3)-glucan and CFW (Life Technologies) to visualize chitin (Wheeler and Fink, 2006 (link); Victoria et al., 2010 (link)). Microscopy was performed on a Leica confocal microscope (Wetzlar, Gernamy) with a 60 × oil objective.