Maximizing the biochemical resolving power of fluorescence microscopy.

Most recent advances in fluorescence microscopy have focused on achieving spatial resolutions below the diffraction limit. However, the inherent capability of fluorescence microscopy to non-invasively resolve different biochemical or physical environments in biological samples has not yet been forma...

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Autores principales:	Alessandro Esposito, Marina Popleteeva, Ashok R Venkitaraman
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ff54720a528c4cf6996a8f2a0f3523b1
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spelling	oai:doaj.org-article:ff54720a528c4cf6996a8f2a0f3523b12021-11-18T08:49:17ZMaximizing the biochemical resolving power of fluorescence microscopy.1932-620310.1371/journal.pone.0077392https://doaj.org/article/ff54720a528c4cf6996a8f2a0f3523b12013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24204821/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Most recent advances in fluorescence microscopy have focused on achieving spatial resolutions below the diffraction limit. However, the inherent capability of fluorescence microscopy to non-invasively resolve different biochemical or physical environments in biological samples has not yet been formally described, because an adequate and general theoretical framework is lacking. Here, we develop a mathematical characterization of the biochemical resolution in fluorescence detection with Fisher information analysis. To improve the precision and the resolution of quantitative imaging methods, we demonstrate strategies for the optimization of fluorescence lifetime, fluorescence anisotropy and hyperspectral detection, as well as different multi-dimensional techniques. We describe optimized imaging protocols, provide optimization algorithms and describe precision and resolving power in biochemical imaging thanks to the analysis of the general properties of Fisher information in fluorescence detection. These strategies enable the optimal use of the information content available within the limited photon-budget typically available in fluorescence microscopy. This theoretical foundation leads to a generalized strategy for the optimization of multi-dimensional optical detection, and demonstrates how the parallel detection of all properties of fluorescence can maximize the biochemical resolving power of fluorescence microscopy, an approach we term Hyper Dimensional Imaging Microscopy (HDIM). Our work provides a theoretical framework for the description of the biochemical resolution in fluorescence microscopy, irrespective of spatial resolution, and for the development of a new class of microscopes that exploit multi-parametric detection systems.Alessandro EspositoMarina PopleteevaAshok R VenkitaramanPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 10, p e77392 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Alessandro Esposito Marina Popleteeva Ashok R Venkitaraman Maximizing the biochemical resolving power of fluorescence microscopy.
description	Most recent advances in fluorescence microscopy have focused on achieving spatial resolutions below the diffraction limit. However, the inherent capability of fluorescence microscopy to non-invasively resolve different biochemical or physical environments in biological samples has not yet been formally described, because an adequate and general theoretical framework is lacking. Here, we develop a mathematical characterization of the biochemical resolution in fluorescence detection with Fisher information analysis. To improve the precision and the resolution of quantitative imaging methods, we demonstrate strategies for the optimization of fluorescence lifetime, fluorescence anisotropy and hyperspectral detection, as well as different multi-dimensional techniques. We describe optimized imaging protocols, provide optimization algorithms and describe precision and resolving power in biochemical imaging thanks to the analysis of the general properties of Fisher information in fluorescence detection. These strategies enable the optimal use of the information content available within the limited photon-budget typically available in fluorescence microscopy. This theoretical foundation leads to a generalized strategy for the optimization of multi-dimensional optical detection, and demonstrates how the parallel detection of all properties of fluorescence can maximize the biochemical resolving power of fluorescence microscopy, an approach we term Hyper Dimensional Imaging Microscopy (HDIM). Our work provides a theoretical framework for the description of the biochemical resolution in fluorescence microscopy, irrespective of spatial resolution, and for the development of a new class of microscopes that exploit multi-parametric detection systems.
format	article
author	Alessandro Esposito Marina Popleteeva Ashok R Venkitaraman
author_facet	Alessandro Esposito Marina Popleteeva Ashok R Venkitaraman
author_sort	Alessandro Esposito
title	Maximizing the biochemical resolving power of fluorescence microscopy.
title_short	Maximizing the biochemical resolving power of fluorescence microscopy.
title_full	Maximizing the biochemical resolving power of fluorescence microscopy.
title_fullStr	Maximizing the biochemical resolving power of fluorescence microscopy.
title_full_unstemmed	Maximizing the biochemical resolving power of fluorescence microscopy.
title_sort	maximizing the biochemical resolving power of fluorescence microscopy.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/ff54720a528c4cf6996a8f2a0f3523b1
work_keys_str_mv	AT alessandroesposito maximizingthebiochemicalresolvingpoweroffluorescencemicroscopy AT marinapopleteeva maximizingthebiochemicalresolvingpoweroffluorescencemicroscopy AT ashokrvenkitaraman maximizingthebiochemicalresolvingpoweroffluorescencemicroscopy
_version_	1718421301935210496

Maximizing the biochemical resolving power of fluorescence microscopy.

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