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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Society of Photo-Optical Instrumentation Engineers
2020
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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. |
format | Online Article Text |
id | pubmed-7008597 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Society of Photo-Optical Instrumentation Engineers |
record_format | MEDLINE/PubMed |
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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