Size photometry and fluorescence imaging of immobilized immersed extracellular vesicles.

ABSTRACT: Immunofluorescence analysis of individual extracellular vesicles (EVs) in common fluorescence microscopes is gaining popularity due to its accessibility and high fluorescence sensitivity; however, EV number and size are only measurable using fluorescent stains requiring extensive sample manipulations. Here we introduce highly sensitive label-free EV size photometry (SP) based on interferometric scattering (iSCAT) imaging of immersed EVs immobilized on a glass coverslip. We implement SP on a common inverted epifluorescence microscope with LED illumination and a simple 50:50 beamsplitter, permitting seamless integration of SP with fluorescence imaging (SPFI). We present a high-throughput SPFI workflow recording >10,000 EVs in 7 min over ten 88 × 88 µm² fields of view, pre- and pos

SUBMITTER: Wallucks A 

PROVIDER: S-EPMC11472239 | biostudies-literature | 2024 Oct

REPOSITORIES: biostudies-literature