Metabolic fingerprinting of bacteria by fluorescence lifetime imaging microscopy

Abstract Bacterial populations exhibit a range of metabolic states influenced by their environment, intra- and interspecies interactions. The identification of bacterial metabolic states and transitions between them in their native environment promises to elucidate community behavior and stochastic...

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Autores principales:	Arunima Bhattacharjee, Rupsa Datta, Enrico Gratton, Allon I. Hochbaum
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/087a549f5bbe4ca3b0cb8e42508fc0c6
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spelling	oai:doaj.org-article:087a549f5bbe4ca3b0cb8e42508fc0c62021-12-02T12:32:34ZMetabolic fingerprinting of bacteria by fluorescence lifetime imaging microscopy10.1038/s41598-017-04032-w2045-2322https://doaj.org/article/087a549f5bbe4ca3b0cb8e42508fc0c62017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04032-whttps://doaj.org/toc/2045-2322Abstract Bacterial populations exhibit a range of metabolic states influenced by their environment, intra- and interspecies interactions. The identification of bacterial metabolic states and transitions between them in their native environment promises to elucidate community behavior and stochastic processes, such as antibiotic resistance acquisition. In this work, we employ two-photon fluorescence lifetime imaging microscopy (FLIM) to create a metabolic fingerprint of individual bacteria and populations. FLIM of autofluorescent reduced nicotinamide adenine dinucleotide (phosphate), NAD(P)H, has been previously exploited for label-free metabolic imaging of mammalian cells. However, NAD(P)H FLIM has not been established as a metabolic proxy in bacteria. Applying the phasor approach, we create FLIM-phasor maps of Escherichia coli, Salmonella enterica serovar Typhimurium, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus epidermidis at the single cell and population levels. The bacterial phasor is sensitive to environmental conditions such as antibiotic exposure and growth phase, suggesting that observed shifts in the phasor are representative of metabolic changes within the cells. The FLIM-phasor approach represents a powerful, non-invasive imaging technique to study bacterial metabolism in situ and could provide unique insights into bacterial community behavior, pathology and antibiotic resistance with sub-cellular resolution.Arunima BhattacharjeeRupsa DattaEnrico GrattonAllon I. HochbaumNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Arunima Bhattacharjee Rupsa Datta Enrico Gratton Allon I. Hochbaum Metabolic fingerprinting of bacteria by fluorescence lifetime imaging microscopy
description	Abstract Bacterial populations exhibit a range of metabolic states influenced by their environment, intra- and interspecies interactions. The identification of bacterial metabolic states and transitions between them in their native environment promises to elucidate community behavior and stochastic processes, such as antibiotic resistance acquisition. In this work, we employ two-photon fluorescence lifetime imaging microscopy (FLIM) to create a metabolic fingerprint of individual bacteria and populations. FLIM of autofluorescent reduced nicotinamide adenine dinucleotide (phosphate), NAD(P)H, has been previously exploited for label-free metabolic imaging of mammalian cells. However, NAD(P)H FLIM has not been established as a metabolic proxy in bacteria. Applying the phasor approach, we create FLIM-phasor maps of Escherichia coli, Salmonella enterica serovar Typhimurium, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus epidermidis at the single cell and population levels. The bacterial phasor is sensitive to environmental conditions such as antibiotic exposure and growth phase, suggesting that observed shifts in the phasor are representative of metabolic changes within the cells. The FLIM-phasor approach represents a powerful, non-invasive imaging technique to study bacterial metabolism in situ and could provide unique insights into bacterial community behavior, pathology and antibiotic resistance with sub-cellular resolution.
format	article
author	Arunima Bhattacharjee Rupsa Datta Enrico Gratton Allon I. Hochbaum
author_facet	Arunima Bhattacharjee Rupsa Datta Enrico Gratton Allon I. Hochbaum
author_sort	Arunima Bhattacharjee
title	Metabolic fingerprinting of bacteria by fluorescence lifetime imaging microscopy
title_short	Metabolic fingerprinting of bacteria by fluorescence lifetime imaging microscopy
title_full	Metabolic fingerprinting of bacteria by fluorescence lifetime imaging microscopy
title_fullStr	Metabolic fingerprinting of bacteria by fluorescence lifetime imaging microscopy
title_full_unstemmed	Metabolic fingerprinting of bacteria by fluorescence lifetime imaging microscopy
title_sort	metabolic fingerprinting of bacteria by fluorescence lifetime imaging microscopy
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/087a549f5bbe4ca3b0cb8e42508fc0c6
work_keys_str_mv	AT arunimabhattacharjee metabolicfingerprintingofbacteriabyfluorescencelifetimeimagingmicroscopy AT rupsadatta metabolicfingerprintingofbacteriabyfluorescencelifetimeimagingmicroscopy AT enricogratton metabolicfingerprintingofbacteriabyfluorescencelifetimeimagingmicroscopy AT allonihochbaum metabolicfingerprintingofbacteriabyfluorescencelifetimeimagingmicroscopy
_version_	1718394018677653504

Metabolic fingerprinting of bacteria by fluorescence lifetime imaging microscopy

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