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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

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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Related Experiment Video

Updated: Jun 2, 2026

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
09:16

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

Published on: January 7, 2019

Structural characterization of individual vesicles using fluorescence microscopy.

Emily C Heider1, Moussa Barhoum, Kyle Edwards

  • 1Department of Chemistry, University of Utah, 315 South 1400 East, Salt Lake City, Utah 84112-0850, USA.

Analytical Chemistry
|May 18, 2011
PubMed
Summary
This summary is machine-generated.

This study introduces a quantitative fluorescence imaging method to analyze individual extruded vesicles, revealing their size, lamellarity, and structural heterogeneity. This technique offers a more detailed understanding of vesicle populations compared to bulk analysis.

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Last Updated: Jun 2, 2026

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
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Direct Stochastic Optical Reconstruction Microscopy of Extracellular Vesicles in Three Dimensions

Published on: August 26, 2021

Area of Science:

  • Lipid vesicle characterization
  • Quantitative fluorescence microscopy
  • Biomembrane structure analysis

Background:

  • Extrusion is a common method for producing uniform lipid vesicles, often assumed to be unilamellar.
  • Bulk analysis methods provide averaged vesicle properties, masking population heterogeneities.
  • Characterizing individual vesicle properties is crucial for understanding their behavior and applications.

Purpose of the Study:

  • To develop and validate a quantitative fluorescence image analysis method for individual extruded vesicles.
  • To determine the size, lamellarity, and structure of vesicles produced by extrusion.
  • To characterize heterogeneities within vesicle populations.

Main Methods:

  • Immobilization of phosphatidylcholine vesicles on biotin-modified glass coverslips via biotin-avidin-biotin binding.
  • Quantitative fluorescence imaging of vesicles containing biotin-modified and NBD-labeled phospholipids.
  • Application of a membrane-impermeable quencher to differentiate outer and inner vesicle layers.
  • Analysis of fluorescence intensity changes to quantify vesicle lamellarity and structure.

Main Results:

  • The developed method allows for the quantitative analysis of individual vesicle size, lamellarity, and structure.
  • Fluorescence microscopy revealed heterogeneities in vesicle populations produced by extrusion.
  • Quantification of bilayer number and structure in individual vesicles was achieved through fractional intensity loss.

Conclusions:

  • Quantitative fluorescence image analysis provides detailed insights into individual vesicle properties, surpassing bulk analysis.
  • Extruded vesicles exhibit a distribution of size, lamellarity, and structure, challenging the assumption of uniform unilamellarity.
  • This method enables precise characterization of vesicle populations for research and development.