Background Reduction in STED-FCS Using a Bivortex Phase Mask.

ACS Photonics
Aurélien BarbotinMartin J Booth

Abstract

Fluorescence correlation spectroscopy (FCS) is a valuable tool to study the molecular dynamics in living cells. When used together with a super-resolution stimulated emission depletion (STED) microscope, STED-FCS can measure diffusion processes on the nanoscale in living cells. In two-dimensional (2D) systems like the cellular plasma membrane, a ring-shaped depletion focus is most commonly used to increase the lateral resolution, leading to more than 25-fold decrease in the observation volume, reaching the relevant scale of supramolecular arrangements. However, STED-FCS faces severe limitations when measuring diffusion in three dimensions (3D), largely due to the spurious background contributions from undepleted areas of the excitation focus that reduce the signal quality and ultimately limit the resolution. In this paper, we investigate how different STED confinement modes can mitigate this issue. By simulations as well as experiments with fluorescent probes in solution and in cells, we demonstrate that the coherent-hybrid (CH) depletion pattern created by a bivortex phase mask reduces background most efficiently and thus provides superior signal quality under comparable reduction of the observation volume. Featuring also the ...Continue Reading

References

Sep 2, 2004·Biophysical Journal·Matthias WeissTommy Nilsson
May 21, 2005·Physical Review Letters·Lars KastrupStefan W Hell
Oct 8, 2014·Annual Review of Cell and Developmental Biology·Anthony A HymanFrank Jülicher
Jul 1, 2015·Methods : a Companion to Methods in Enzymology·Mathias P ClausenChristian Eggeling
Jul 30, 2016·Optics Letters·Jacopo AntonelloMartin J Booth
Jul 8, 2017·Nature Communications·Luca LanzanòGiuseppe Vicidomini
Sep 1, 2017·Biomedical Optics Express·Raphaël TurcotteNa Ji
Jun 5, 2018·Optics Communications·Jacopo AntonelloMartin J Booth
Jan 10, 2019·Nanoscale·Giorgio TortaroloGiuseppe Vicidomini
Mar 8, 2019·Nature Protocols·Erdinc SezginChristian Eggeling
Aug 21, 2019·Annual Review of Genetics·Simon Alberti, Dorothee Dormann
Sep 13, 2019·Optics Express·Aurélien BarbotinMartin J Booth
Oct 9, 2019·Genes & Development·David T McSwiggenRobert Tjian
Nov 17, 2020·Journal of Physics D: Applied Physics·Falk SchneiderIztok Urbančič

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Citations

Jul 10, 2021·Optica·Mary Grace M VelascoJoerg Bewersdorf

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Methods Mentioned

BETA
FCS

Software Mentioned

STED
SPLIT
multipletau
FCS
STEDD
CH

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