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Single-shot time-folded fluorescence lifetime imaging

Fluorescence lifetime imaging is an important tool in bioimaging that allows one to detect subtle changes in cell dynamics and their environment. Most time-domain approaches currently involve scanning a single illumination point across the sample, which can make imaging dynamic scenes challenging, w...

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Autores principales: Kapitany, Valentin, Zickus, Vytautas, Fatima, Areeba, Carles, Guillem, Faccio, Daniele
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10120087/
https://www.ncbi.nlm.nih.gov/pubmed/37043531
http://dx.doi.org/10.1073/pnas.2214617120
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author Kapitany, Valentin
Zickus, Vytautas
Fatima, Areeba
Carles, Guillem
Faccio, Daniele
author_facet Kapitany, Valentin
Zickus, Vytautas
Fatima, Areeba
Carles, Guillem
Faccio, Daniele
author_sort Kapitany, Valentin
collection PubMed
description Fluorescence lifetime imaging is an important tool in bioimaging that allows one to detect subtle changes in cell dynamics and their environment. Most time-domain approaches currently involve scanning a single illumination point across the sample, which can make imaging dynamic scenes challenging, while single-shot “rapid lifetime determination” can suffer from large uncertainties when the lifetime is not appropriately sampled. Here, we propose a time-folded fluorescence lifetime imaging microscopy (TFFLIM) approach, whereby a time-folding cavity provides multiple spatially sheared replicas of the lifetime, each shifted temporally with respect to a fixed time gate. This provides a robust, single-shot FLIM approach that we experimentally validate across a broad lifetime range on fluorescent beads and Convallaria samples.
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spelling pubmed-101200872023-04-22 Single-shot time-folded fluorescence lifetime imaging Kapitany, Valentin Zickus, Vytautas Fatima, Areeba Carles, Guillem Faccio, Daniele Proc Natl Acad Sci U S A Biological Sciences Fluorescence lifetime imaging is an important tool in bioimaging that allows one to detect subtle changes in cell dynamics and their environment. Most time-domain approaches currently involve scanning a single illumination point across the sample, which can make imaging dynamic scenes challenging, while single-shot “rapid lifetime determination” can suffer from large uncertainties when the lifetime is not appropriately sampled. Here, we propose a time-folded fluorescence lifetime imaging microscopy (TFFLIM) approach, whereby a time-folding cavity provides multiple spatially sheared replicas of the lifetime, each shifted temporally with respect to a fixed time gate. This provides a robust, single-shot FLIM approach that we experimentally validate across a broad lifetime range on fluorescent beads and Convallaria samples. National Academy of Sciences 2023-04-12 2023-04-18 /pmc/articles/PMC10120087/ /pubmed/37043531 http://dx.doi.org/10.1073/pnas.2214617120 Text en Copyright © 2023 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by/4.0/This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Biological Sciences
Kapitany, Valentin
Zickus, Vytautas
Fatima, Areeba
Carles, Guillem
Faccio, Daniele
Single-shot time-folded fluorescence lifetime imaging
title Single-shot time-folded fluorescence lifetime imaging
title_full Single-shot time-folded fluorescence lifetime imaging
title_fullStr Single-shot time-folded fluorescence lifetime imaging
title_full_unstemmed Single-shot time-folded fluorescence lifetime imaging
title_short Single-shot time-folded fluorescence lifetime imaging
title_sort single-shot time-folded fluorescence lifetime imaging
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10120087/
https://www.ncbi.nlm.nih.gov/pubmed/37043531
http://dx.doi.org/10.1073/pnas.2214617120
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