Quantifying Candida albicans Biofilm Inhibition

First, C. albicans ATCC 60193 inoculum (2.5 × 10⁵ CFU/mL) was added to the wells of 96-well microplates along with CAT at different concentrations (MIC, 2 × MIC and 4 × MIC). The plate was incubated for 2 h at 37 °C to allow for initial adherence. Then the wells were washed with PBS in order to remove unbound cells; fresh SDB was added, and the plates were incubated again for 48 h at 37 °C. For biofilm quantification, the wells were washed twice with PBS, air-dried for 45 min, dyed with 0.4% crystal violet and unstained with 95% EtOH. Absorbance values were read at 600 nm using a microplate reader (GloMax-Multi, PROMEGA, Madison, WI, USA) [40 (link)].