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

STAR Protoc

Instituto de Biofísica Carlos Chagas Filho (IBCCF), Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro 21941-904, Brazil. Electronic address:

Published: June 2024


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Article Abstract

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., Rizzo et al., Reis et al., and Reis et al..

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11135028PMC
http://dx.doi.org/10.1016/j.xpro.2024.103069DOI Listing

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