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Forschungsdatenbank PMU-SQQUID

Differential fluorescence nanoparticle tracking analysis for enumeration of the extracellular vesicle content in mixed particulate solutions.
Desgeorges, A; Hollerweger, J; Lassacher, T; Rohde, E; Helmbrecht, C; Gimona, M
METHODS. 2020; 177: 67-73.
Originalarbeiten (Zeitschrift)

PMU-Autor/inn/en

Desgeorges Alexandre
Gimona Mario
Hollerweger Julia Carolin
Lassacher Teresa
Rohde Eva

Abstract

A major concern for the extracellular vesicle (EV) field is the current lack of accurate methods for EV quantification. Total protein measurement fails to reliably quantify EVs from serum-containing conditioned media and classical nanoparticle tracking analysis (NTA) allows quantification and size determination of particles, but fails to discriminate between membrane-bounded EVs, lipids and protein aggregates. However, EVs can be fluorescently labelled with non-specific membrane markers or with antibodies specifically recognizing EV surface marker proteins. Fluorescence-based NTA (F-NTA) is thus emerging as a method for counting and phenotyping of EVs. We have validated a differential NTA/F-NTA method using specific antibodies against surface markers in analogy to flow cytometric analyses.