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Single molecule confocal fluorescence lifetime correlation spectroscopy for accurate nanoparticle size determination

Published

Author(s)

Bonghwan Chon, Kimberly Briggman, Jeeseong C. Hwang

Abstract

We report on an experimental procedure in confocal single molecule fluorescence lifetime correlation spectroscopy (FLCS) to determine the range of excitation power and molecule concentration in solution under which the application of an unmodified model autocorrelation function is justified. This procedure enables fitting of the autocorrelation to an accurate model to measure diffusion length (r) and diffusion time (τD) of single molecules in solution. We also report on the pinhole size dependency of r and τD in a confocal FLCS platform. This procedure determines a set of experimental parameters with which the Stoke-Einstein equation accurately measures the hydrodynamic radii of spherical nanoparticles, enabling the determination of the particle size range for which the hydrodynamic radius by the S-E equation measures the real particle radius.
Citation
Physical Chemistry Chemical Physics
Volume
16

Keywords

single molecule, Fluorescence correlation spectroscopy, FCS, nanoparticle, fluorescence lifetime, confocal microscopy, Fluorescence lifetime correlation spectroscopy, FLCS

Citation

Chon, B. , Briggman, K. and Hwang, J. (2014), Single molecule confocal fluorescence lifetime correlation spectroscopy for accurate nanoparticle size determination, Physical Chemistry Chemical Physics, [online], https://doi.org/10.1039/C4CP01197J (Accessed March 28, 2024)
Created May 14, 2014, Updated October 12, 2021