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Optical changes in THP-1 macrophage metabolism in response to pro- and anti-inflammatory stimuli reported by label-free two-photon imaging

Temporal changes in macrophage metabolism are likely crucial to their role in inflammatory diseases. Label-free two-photon excited fluorescence (TPEF) and fluorescence lifetime imaging microscopy are well suited to track dynamic changes in macrophage metabolism. We performed TPEF imaging of human ma...

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Autores principales: Smokelin, Isabel S., Mizzoni, Craig, Erndt-Marino, Josh, Kaplan, David L., Georgakoudi, Irene
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Society of Photo-Optical Instrumentation Engineers 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7008597/
https://www.ncbi.nlm.nih.gov/pubmed/31953928
http://dx.doi.org/10.1117/1.JBO.25.1.014512
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author Smokelin, Isabel S.
Mizzoni, Craig
Erndt-Marino, Josh
Kaplan, David L.
Georgakoudi, Irene
author_facet Smokelin, Isabel S.
Mizzoni, Craig
Erndt-Marino, Josh
Kaplan, David L.
Georgakoudi, Irene
author_sort Smokelin, Isabel S.
collection PubMed
description Temporal changes in macrophage metabolism are likely crucial to their role in inflammatory diseases. Label-free two-photon excited fluorescence (TPEF) and fluorescence lifetime imaging microscopy are well suited to track dynamic changes in macrophage metabolism. We performed TPEF imaging of human macrophages following either pro- or an anti-inflammatory stimulation. Two endogenous fluorophores, NAD(P)H and FAD, coenzymes involved in key metabolic pathways, provided contrast. We used the corresponding intensity images to determine the optical redox ratio of FAD to FAD + NAD(P)H. We also analyzed the intensity fluctuation patterns within NAD(P)H TPEF images to determine mitochondrial clustering patterns. Finally, we acquired NAD(P)H TPEF lifetime images to assess the relative levels of bound NAD(P)H. Our studies indicate that the redox ratio increases, whereas mitochondrial clustering decreases in response to both pro- and anti-inflammatory stimuli; however, these changes are enhanced in pro-inflammatory macrophages. Interestingly, we did not detect any significant changes in the corresponding NAD(P)H bound fraction. A combination of optical metabolic metrics could be used to classify pro- and anti-inflammatory macrophages with high accuracy. Contributions from alterations in different metabolic pathways may explain our findings, which highlight the potential of label-free two-photon imaging to assess nondestructively macrophage functional state.
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spelling pubmed-70085972020-02-14 Optical changes in THP-1 macrophage metabolism in response to pro- and anti-inflammatory stimuli reported by label-free two-photon imaging Smokelin, Isabel S. Mizzoni, Craig Erndt-Marino, Josh Kaplan, David L. Georgakoudi, Irene J Biomed Opt Special Section Celebrating Thirty Years of Multiphoton Microscopy in the Biomedical Sciences Temporal changes in macrophage metabolism are likely crucial to their role in inflammatory diseases. Label-free two-photon excited fluorescence (TPEF) and fluorescence lifetime imaging microscopy are well suited to track dynamic changes in macrophage metabolism. We performed TPEF imaging of human macrophages following either pro- or an anti-inflammatory stimulation. Two endogenous fluorophores, NAD(P)H and FAD, coenzymes involved in key metabolic pathways, provided contrast. We used the corresponding intensity images to determine the optical redox ratio of FAD to FAD + NAD(P)H. We also analyzed the intensity fluctuation patterns within NAD(P)H TPEF images to determine mitochondrial clustering patterns. Finally, we acquired NAD(P)H TPEF lifetime images to assess the relative levels of bound NAD(P)H. Our studies indicate that the redox ratio increases, whereas mitochondrial clustering decreases in response to both pro- and anti-inflammatory stimuli; however, these changes are enhanced in pro-inflammatory macrophages. Interestingly, we did not detect any significant changes in the corresponding NAD(P)H bound fraction. A combination of optical metabolic metrics could be used to classify pro- and anti-inflammatory macrophages with high accuracy. Contributions from alterations in different metabolic pathways may explain our findings, which highlight the potential of label-free two-photon imaging to assess nondestructively macrophage functional state. Society of Photo-Optical Instrumentation Engineers 2020-01-17 2020-01 /pmc/articles/PMC7008597/ /pubmed/31953928 http://dx.doi.org/10.1117/1.JBO.25.1.014512 Text en © 2020 The Authors https://creativecommons.org/licenses/by/4.0/ Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
spellingShingle Special Section Celebrating Thirty Years of Multiphoton Microscopy in the Biomedical Sciences
Smokelin, Isabel S.
Mizzoni, Craig
Erndt-Marino, Josh
Kaplan, David L.
Georgakoudi, Irene
Optical changes in THP-1 macrophage metabolism in response to pro- and anti-inflammatory stimuli reported by label-free two-photon imaging
title Optical changes in THP-1 macrophage metabolism in response to pro- and anti-inflammatory stimuli reported by label-free two-photon imaging
title_full Optical changes in THP-1 macrophage metabolism in response to pro- and anti-inflammatory stimuli reported by label-free two-photon imaging
title_fullStr Optical changes in THP-1 macrophage metabolism in response to pro- and anti-inflammatory stimuli reported by label-free two-photon imaging
title_full_unstemmed Optical changes in THP-1 macrophage metabolism in response to pro- and anti-inflammatory stimuli reported by label-free two-photon imaging
title_short Optical changes in THP-1 macrophage metabolism in response to pro- and anti-inflammatory stimuli reported by label-free two-photon imaging
title_sort optical changes in thp-1 macrophage metabolism in response to pro- and anti-inflammatory stimuli reported by label-free two-photon imaging
topic Special Section Celebrating Thirty Years of Multiphoton Microscopy in the Biomedical Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7008597/
https://www.ncbi.nlm.nih.gov/pubmed/31953928
http://dx.doi.org/10.1117/1.JBO.25.1.014512
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