1 Maintain C. albicans by weekly transfer on Sabouraud dextrose agar plates at 28°C.
2 Transfer a colony from a three-days-old culture to 5 ml PBS in 15 ml polypropylene tube using a sterile loop.
3 Wash twice with PBS by centrifugation at 200 g for 10 min.
4 Resuspend the yeasts at 4 x 10s/ml in complete RPMI medium,
5 Plate 0.1 ml (5 x lO4 yeasts} per well in 96-weIl, flat-bottom microtitre plates with a repeating dispenser.
6 Incubate plates for 3-6 h at 37 °C until 95% of Candida have germinated into hyphae (30-100 ^m in length) that adhere firmly to the bottom of the wells (check hyphae germination by inverted microscope examination).3
7 Remove supernatant from each well by pipette aspiration observing biohazard precautions.
Was this article helpful?