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Fluorescence correlation spectroscopy for particle sizing: A notorious challenge.

Jan-Hagen Krohn1, Adam Mamot2, Nastasja Kaletta2

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Fluorescence correlation spectroscopy (FCS) offers precise particle sizing but faces challenges with complex samples. This review details strategies to overcome these hurdles for routine biophysical applications.

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Area of Science:

  • Biophysics
  • Nanotechnology
  • Materials Science

Background:

  • Accurate particle size determination is crucial for biological systems.
  • Fluorescence correlation spectroscopy (FCS) offers high sensitivity for sizing small particles.
  • FCS provides advantages over dynamic light scattering (DLS) due to fluorescence detection.

Purpose of the Study:

  • To discuss systematic challenges in applying FCS for particle sizing in polydisperse samples.
  • To review advances in FCS methodology and data analysis for improved accuracy.
  • To guide users in overcoming limitations for routine FCS application.

Main Methods:

  • Review of existing literature on FCS for particle sizing.
  • Analysis of challenges related to polydispersity and fluorescent labeling.
  • Discussion of error mitigation strategies in experimental design and data analysis.

Main Results:

  • FCS sensitivity, while advantageous, can lead to errors in complex samples.
  • Large particles and fluorescent labeling effects pose significant challenges in FCS analysis.
  • Strategies exist to mitigate errors and improve the reliability of FCS measurements.

Conclusions:

  • Despite challenges, advances enable more robust FCS application for particle sizing.
  • Addressing issues like polydispersity and labeling is key to wider FCS adoption.
  • This work aims to foster routine use of FCS in biophysical research.