Back to the Basics: Two Approaches for the Identification and Extraction of Lipid Droplets from <i>Malassezia pachydermatis</i> CBS1879 and <i>Malassezia globosa</i> CBS7966

Malassezia spp. are lipid-dependent yeasts that have been related to skin mycobiota and dermatological systemic diseases. Study of lipid droplets (LDs) is relevant elucidate the unknown role these organelles in gain a broader overview metabolism Malassezia. Here, we standardized two protocols for analysis LDs M. pachydermatis globosa. The first describes co-staining confocal laser-scanning fluorescence microscopy, second details extraction purification LDs. double stain achieved with three different neutral fluorophores, namely Nile Red, BODIPY™ 493/503, HCS LipidTOX™ Deep Red Neutral, combination Calcofluor White. For LD extraction, cell wall rupture conducted using Trichoderma harzianum enzymes cycles vortexing zirconium beads. performed three-step ultracentrifugation process. These standardizations will contribute study dynamics, morphology, composition © 2021 Wiley Periodicals LLC. Basic Protocol 1: Lipid droplet staining 2: Support Protocol: culture conditions