Advanced Data Analysis for Fluorescence-Lifetime Single-Molecule Localization Microscopy

Fluorescence-lifetime single molecule localization microscopy (FL-SMLM) adds the lifetime dimension to the spatial super-resolution provided by SMLM. Independent of intensity and spectrum, this lifetime information can be used, for example, to quantify the energy transfer efficiency in Förster Reson...

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Autores principales:	Jan Christoph Thiele, Oleksii Nevskyi, Dominic A. Helmerich, Markus Sauer, Jörg Enderlein
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	FLIM (fluorescence lifetime imaging microscopy) CRLB (Cramér-Rao lower bound) analysis fluoresence lifetime fitting super-resolution microscopy lifetime uncertainty SMLM (single molecule localisation microscopy) Computer applications to medicine. Medical informatics R858-859.7
Acceso en línea:	https://doaj.org/article/84e99b9e96e44937a66bb425f84b4dcd
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spelling	oai:doaj.org-article:84e99b9e96e44937a66bb425f84b4dcd2021-11-19T05:39:53ZAdvanced Data Analysis for Fluorescence-Lifetime Single-Molecule Localization Microscopy2673-764710.3389/fbinf.2021.740281https://doaj.org/article/84e99b9e96e44937a66bb425f84b4dcd2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fbinf.2021.740281/fullhttps://doaj.org/toc/2673-7647Fluorescence-lifetime single molecule localization microscopy (FL-SMLM) adds the lifetime dimension to the spatial super-resolution provided by SMLM. Independent of intensity and spectrum, this lifetime information can be used, for example, to quantify the energy transfer efficiency in Förster Resonance Energy Transfer (FRET) imaging, to probe the local environment with dyes that change their lifetime in an environment-sensitive manner, or to achieve image multiplexing by using dyes with different lifetimes. We present a thorough theoretical analysis of fluorescence-lifetime determination in the context of FL-SMLM and compare different lifetime-fitting approaches. In particular, we investigate the impact of background and noise, and give clear guidelines for procedures that are optimized for FL-SMLM. We do also present and discuss our public-domain software package “Fluorescence-Lifetime TrackNTrace,” which converts recorded fluorescence microscopy movies into super-resolved FL-SMLM images.Jan Christoph ThieleOleksii NevskyiDominic A. HelmerichMarkus SauerJörg EnderleinJörg EnderleinFrontiers Media S.A.articleFLIM (fluorescence lifetime imaging microscopy)CRLB (Cramér-Rao lower bound) analysisfluoresence lifetime fittingsuper-resolution microscopylifetime uncertaintySMLM (single molecule localisation microscopy)Computer applications to medicine. Medical informaticsR858-859.7ENFrontiers in Bioinformatics, Vol 1 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	FLIM (fluorescence lifetime imaging microscopy) CRLB (Cramér-Rao lower bound) analysis fluoresence lifetime fitting super-resolution microscopy lifetime uncertainty SMLM (single molecule localisation microscopy) Computer applications to medicine. Medical informatics R858-859.7
spellingShingle	FLIM (fluorescence lifetime imaging microscopy) CRLB (Cramér-Rao lower bound) analysis fluoresence lifetime fitting super-resolution microscopy lifetime uncertainty SMLM (single molecule localisation microscopy) Computer applications to medicine. Medical informatics R858-859.7 Jan Christoph Thiele Oleksii Nevskyi Dominic A. Helmerich Markus Sauer Jörg Enderlein Jörg Enderlein Advanced Data Analysis for Fluorescence-Lifetime Single-Molecule Localization Microscopy
description	Fluorescence-lifetime single molecule localization microscopy (FL-SMLM) adds the lifetime dimension to the spatial super-resolution provided by SMLM. Independent of intensity and spectrum, this lifetime information can be used, for example, to quantify the energy transfer efficiency in Förster Resonance Energy Transfer (FRET) imaging, to probe the local environment with dyes that change their lifetime in an environment-sensitive manner, or to achieve image multiplexing by using dyes with different lifetimes. We present a thorough theoretical analysis of fluorescence-lifetime determination in the context of FL-SMLM and compare different lifetime-fitting approaches. In particular, we investigate the impact of background and noise, and give clear guidelines for procedures that are optimized for FL-SMLM. We do also present and discuss our public-domain software package “Fluorescence-Lifetime TrackNTrace,” which converts recorded fluorescence microscopy movies into super-resolved FL-SMLM images.
format	article
author	Jan Christoph Thiele Oleksii Nevskyi Dominic A. Helmerich Markus Sauer Jörg Enderlein Jörg Enderlein
author_facet	Jan Christoph Thiele Oleksii Nevskyi Dominic A. Helmerich Markus Sauer Jörg Enderlein Jörg Enderlein
author_sort	Jan Christoph Thiele
title	Advanced Data Analysis for Fluorescence-Lifetime Single-Molecule Localization Microscopy
title_short	Advanced Data Analysis for Fluorescence-Lifetime Single-Molecule Localization Microscopy
title_full	Advanced Data Analysis for Fluorescence-Lifetime Single-Molecule Localization Microscopy
title_fullStr	Advanced Data Analysis for Fluorescence-Lifetime Single-Molecule Localization Microscopy
title_full_unstemmed	Advanced Data Analysis for Fluorescence-Lifetime Single-Molecule Localization Microscopy
title_sort	advanced data analysis for fluorescence-lifetime single-molecule localization microscopy
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/84e99b9e96e44937a66bb425f84b4dcd
work_keys_str_mv	AT janchristophthiele advanceddataanalysisforfluorescencelifetimesinglemoleculelocalizationmicroscopy AT oleksiinevskyi advanceddataanalysisforfluorescencelifetimesinglemoleculelocalizationmicroscopy AT dominicahelmerich advanceddataanalysisforfluorescencelifetimesinglemoleculelocalizationmicroscopy AT markussauer advanceddataanalysisforfluorescencelifetimesinglemoleculelocalizationmicroscopy AT jorgenderlein advanceddataanalysisforfluorescencelifetimesinglemoleculelocalizationmicroscopy AT jorgenderlein advanceddataanalysisforfluorescencelifetimesinglemoleculelocalizationmicroscopy
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Advanced Data Analysis for Fluorescence-Lifetime Single-Molecule Localization Microscopy

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