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Super-resolution Fluorescence Microscopy

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.

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Updated: Jun 12, 2026

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis

Published on: January 7, 2019

Fluorescence Characterization of Extracellular Vesicles Using Single-Molecule Confocal Microscopy.

Tianxiao Zhao^1,2, Noelia Pelegrina-Hidalgo^1,2, Daniel C Edwards^1,2

¹EaStCHEM School of Chemistry, The University of Edinburgh, Edinburgh, EH9 3FJ, UK.

|August 18, 2025

Summary

This summary is machine-generated.

This study presents a new method for detecting and quantifying extracellular vesicles (EVs) using fluorescently labeled antibodies and advanced microscopy. The technique achieves high sensitivity, enabling EV biomarker discovery in biological samples.

Keywords:

EVs fluorescence microscopy proteins single‐molecule

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Last Updated: Jun 12, 2026

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis

Published on: January 7, 2019

Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis

Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis

Published on: February 14, 2022

Single Extracellular Vesicle Transmembrane Protein Characterization by Nano-Flow Cytometry

Single Extracellular Vesicle Transmembrane Protein Characterization by Nano-Flow Cytometry

Published on: July 26, 2022

Area of Science:

Biotechnology
Cell Biology
Biomarker Discovery

Background:

Extracellular vesicles (EVs) are crucial for intercellular communication and are emerging as significant biomarkers.
Conventional methods struggle with the low abundance and heterogeneity of EVs, hindering accurate characterization.
Specific detection of individual EVs is essential for reliable biomarker analysis.

Purpose of the Study:

To develop a highly sensitive and specific method for detecting and quantifying individual extracellular vesicles (EVs).
To enable the characterization of EVs directly in complex biological fluids like serum and plasma.
To compare EV extraction yields from different purification methods.

Main Methods:

Coupling EV-specific antibodies with two different fluorophores.
Utilizing fast-flow microfluidics and single-molecule confocal microscopy for EV detection.
Comparing ultracentrifugation and size exclusion chromatography for EV yield.

Main Results:

Achieved sub-femtomolar detection limits (≈10^7 EVs mL^-1) for individual EVs.
Demonstrated specific detection of EVs amidst other lipid vesicles.
Quantified EVs in serum and plasma without prior purification.
Compared EV yields from different isolation techniques.

Conclusions:

The developed method offers a highly specific, sensitive, and user-friendly solution for EV characterization.
This approach facilitates robust EV biomarker discovery and quantification in clinical samples.
The technique overcomes limitations of conventional methods for analyzing heterogeneous EV populations.