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© Institut Pasteur
Cryptococcus neoformans
Publication : STAR protocols

Protocol for separation of fungal extracellular vesicles using ultracentrifugation from solid medium cultures.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in STAR protocols - 20 May 2024

Piffer AC, Reis FCG, Moyrand F, Montalvão B, Rodrigues ML, Janbon G, Rizzo J

Link to Pubmed [PMID] – 38771694

Link to DOI – 10.1016/j.xpro.2024.103069

STAR Protoc 2024 May; 5(2): 103069

Extracellular vesicles (EVs) have been identified in diverse fungi, including human pathogens. In this protocol, we present two techniques for isolating and analyzing fungal EVs. The first is for high-throughput screening, and the second is for yielding concentrated samples suitable for centrifugation-based density gradients. We describe steps for analytical assays such as nano-flow cytometry and nanoparticle tracking analysis to measure EV dimensions and concentration. EV suspensions can serve diverse assays, including electron microscopy, compositional determination, and cell-to-cell communication assays. For complete details on the use and execution of this protocol, please refer to Rizzo et al.,1 Rizzo et al.,2 Reis et al.,3 and Reis et al.4.