Calcofluor white staining

A glycerol stock of DBY10148 was streaked out onto a YPD plate, and a single colony was used to inoculate 30 mL of synthetic complete media. The yeast culture was allowed to grow at 37 °C for 16 hours with shaking at 250 rpm. 100 μL of the late log-phase culture (OD600 = 2.85) was centrifuged at 180 x g for 1 minute to pellet the yeast and resuspended in 40 uL of a solution containing a 1:1 ratio of calcofluor white staining solution (Sigma, Cat. # 18909) and 10% KOH (w/v) as per manufacturer’s instructions. The yeast suspension was incubated in the dark for 1 minute before transferring onto a microscope slide and imaging. Yeast were imaged using a Nikon Eclipse Ti2 with 60x, 1.4 N.A. objective. Images were obtained by a Hamamatsu CMOS camera controlled by NIS-Elements BR 413.04 64-bit software (Nikon). Data were further processed using FIJI (National Institutes of Health).