Time- and spectrally resolved characteristics of flavin fluorescence in U87MG cancer cells in culture
Early detection of cancer is crucial for the successful diagnostics of its presence and its subsequent treatment. To improve cancer detection, we tested the progressive multimodal optical imaging of U87MG cells in culture. A combination of steady-state spectroscopic methods with the time-resolved ap...
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Veröffentlicht in: | Journal of biomedical optics 2015-05, Vol.20 (5), p.051017-051017 |
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description | Early detection of cancer is crucial for the successful diagnostics of its presence and its subsequent treatment. To improve cancer detection, we tested the progressive multimodal optical imaging of U87MG cells in culture. A combination of steady-state spectroscopic methods with the time-resolved approach provides a new insight into the native metabolism when focused on endogenous tissue fluorescence. In this contribution, we evaluated the metabolic state of living U87MG cancer cells in culture by means of endogenous flavin fluorescence. Confocal microscopy and time-resolved fluorescence imaging were employed to gather spectrally and time-resolved images of the flavin fluorescence. We observed that flavin fluorescence in U87MG cells was predominantly localized outside the cell nucleus in mitochondria, while exhibiting a spectral maximum under 500 nm and fluorescence lifetimes under 1.4 ns, suggesting the presence of bound flavins. In some cells, flavin fluorescence was also detected inside the cell nuclei in the nucleoli, exhibiting longer fluorescence lifetimes and a red-shifted spectral maximum, pointing to the presence of free flavin. Extra-nuclear flavin fluorescence was diminished by 2-deoxyglucose, but failed to increase with 2,4-dinitrophenol, the uncoupler of oxidative phosphorylation, indicating that the cells use glycolysis, rather than oxidative phosphorylation for functioning. These gathered data are the first step toward monitoring the metabolic state of U87MG cancer cells. |
doi_str_mv | 10.1117/1.JBO.20.5.051017 |
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To improve cancer detection, we tested the progressive multimodal optical imaging of U87MG cells in culture. A combination of steady-state spectroscopic methods with the time-resolved approach provides a new insight into the native metabolism when focused on endogenous tissue fluorescence. In this contribution, we evaluated the metabolic state of living U87MG cancer cells in culture by means of endogenous flavin fluorescence. Confocal microscopy and time-resolved fluorescence imaging were employed to gather spectrally and time-resolved images of the flavin fluorescence. We observed that flavin fluorescence in U87MG cells was predominantly localized outside the cell nucleus in mitochondria, while exhibiting a spectral maximum under 500 nm and fluorescence lifetimes under 1.4 ns, suggesting the presence of bound flavins. In some cells, flavin fluorescence was also detected inside the cell nuclei in the nucleoli, exhibiting longer fluorescence lifetimes and a red-shifted spectral maximum, pointing to the presence of free flavin. Extra-nuclear flavin fluorescence was diminished by 2-deoxyglucose, but failed to increase with 2,4-dinitrophenol, the uncoupler of oxidative phosphorylation, indicating that the cells use glycolysis, rather than oxidative phosphorylation for functioning. These gathered data are the first step toward monitoring the metabolic state of U87MG cancer cells.</description><identifier>ISSN: 1083-3668</identifier><identifier>EISSN: 1560-2281</identifier><identifier>DOI: 10.1117/1.JBO.20.5.051017</identifier><identifier>PMID: 25521208</identifier><language>eng</language><publisher>United States: Society of Photo-Optical Instrumentation Engineers</publisher><subject>2,4-Dinitrophenol - chemistry ; Cancer ; Cell Line, Tumor ; Cell Nucleolus - metabolism ; Cell Nucleus - metabolism ; Confocal ; Culture ; Deoxyglucose - chemistry ; Disease Progression ; Early Detection of Cancer - methods ; Flavins - chemistry ; Fluorescence ; Fluorescent Dyes - chemistry ; Glycolysis ; Humans ; Imaging ; Microscopy - methods ; Microscopy, Confocal ; Mitochondria - metabolism ; Monitoring ; Nuclei ; Optics and Photonics - methods ; Oxidative Phosphorylation ; Oxygen - chemistry ; Phosphorylation ; Spectra ; Spectrometry, Fluorescence - instrumentation ; Spectrometry, Fluorescence - methods ; Spectrophotometry</subject><ispartof>Journal of biomedical optics, 2015-05, Vol.20 (5), p.051017-051017</ispartof><rights>2015 Society of Photo-Optical Instrumentation Engineers (SPIE)</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c493t-55eebbf6389fd2d14e0827840794562ffa3b118768b8aafd9c9f031f9111a8f33</citedby><cites>FETCH-LOGICAL-c493t-55eebbf6389fd2d14e0827840794562ffa3b118768b8aafd9c9f031f9111a8f33</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25521208$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Horilova, Julia</creatorcontrib><creatorcontrib>Cunderlikova, Beata</creatorcontrib><creatorcontrib>Marcek Chorvatova, Alzbeta</creatorcontrib><title>Time- and spectrally resolved characteristics of flavin fluorescence in U87MG cancer cells in culture</title><title>Journal of biomedical optics</title><addtitle>J. Biomed. Opt</addtitle><description>Early detection of cancer is crucial for the successful diagnostics of its presence and its subsequent treatment. To improve cancer detection, we tested the progressive multimodal optical imaging of U87MG cells in culture. A combination of steady-state spectroscopic methods with the time-resolved approach provides a new insight into the native metabolism when focused on endogenous tissue fluorescence. In this contribution, we evaluated the metabolic state of living U87MG cancer cells in culture by means of endogenous flavin fluorescence. Confocal microscopy and time-resolved fluorescence imaging were employed to gather spectrally and time-resolved images of the flavin fluorescence. We observed that flavin fluorescence in U87MG cells was predominantly localized outside the cell nucleus in mitochondria, while exhibiting a spectral maximum under 500 nm and fluorescence lifetimes under 1.4 ns, suggesting the presence of bound flavins. In some cells, flavin fluorescence was also detected inside the cell nuclei in the nucleoli, exhibiting longer fluorescence lifetimes and a red-shifted spectral maximum, pointing to the presence of free flavin. Extra-nuclear flavin fluorescence was diminished by 2-deoxyglucose, but failed to increase with 2,4-dinitrophenol, the uncoupler of oxidative phosphorylation, indicating that the cells use glycolysis, rather than oxidative phosphorylation for functioning. These gathered data are the first step toward monitoring the metabolic state of U87MG cancer cells.</description><subject>2,4-Dinitrophenol - chemistry</subject><subject>Cancer</subject><subject>Cell Line, Tumor</subject><subject>Cell Nucleolus - metabolism</subject><subject>Cell Nucleus - metabolism</subject><subject>Confocal</subject><subject>Culture</subject><subject>Deoxyglucose - chemistry</subject><subject>Disease Progression</subject><subject>Early Detection of Cancer - methods</subject><subject>Flavins - chemistry</subject><subject>Fluorescence</subject><subject>Fluorescent Dyes - chemistry</subject><subject>Glycolysis</subject><subject>Humans</subject><subject>Imaging</subject><subject>Microscopy - methods</subject><subject>Microscopy, Confocal</subject><subject>Mitochondria - metabolism</subject><subject>Monitoring</subject><subject>Nuclei</subject><subject>Optics and Photonics - methods</subject><subject>Oxidative Phosphorylation</subject><subject>Oxygen - chemistry</subject><subject>Phosphorylation</subject><subject>Spectra</subject><subject>Spectrometry, Fluorescence - instrumentation</subject><subject>Spectrometry, Fluorescence - methods</subject><subject>Spectrophotometry</subject><issn>1083-3668</issn><issn>1560-2281</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU9v1DAQxS1ERf_AB-CCfOSS4LHXjn0shRZQq_awPVuOMxapvEmwk0rl0-Ptlh6gak_jGf_eyH6PkPfAagBoPkH94_NlzVktayaBQfOKHIBUrOJcw-tyZlpUQim9Tw5zvmGMaWXUG7LPpeTAmT4guO43WFE3dDRP6OfkYryjCfMYb7Gj_qdLzs-Y-jz3PtMx0BDdbT-UsowF8zh4pKW_1s3FGfWutIl6jDFvp36J85LwLdkLLmZ891CPyPXp1_XJt-r88uz7yfF55VdGzJWUiG0blNAmdLyDFTLNG71ijVlJxUNwogXQjdKtdi50xpvABART3HA6CHFEPu72Tmn8tWCe7abP28e4AcclW2hYsYmVv7-MKmFMI4ubBYUd6tOYc8Jgp9RvXLqzwOw2CAu2BGE5s9LugiiaDw_rl3aD3aPir_MFqHdAnnq0N-OShuLMsxvXTwkesd_99K_mfnacSnIRr76c_nc9dUH8AbGVrLk</recordid><startdate>20150501</startdate><enddate>20150501</enddate><creator>Horilova, Julia</creator><creator>Cunderlikova, Beata</creator><creator>Marcek Chorvatova, Alzbeta</creator><general>Society of Photo-Optical Instrumentation Engineers</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7SP</scope><scope>7U5</scope><scope>8FD</scope><scope>F28</scope><scope>FR3</scope><scope>L7M</scope></search><sort><creationdate>20150501</creationdate><title>Time- and spectrally resolved characteristics of flavin fluorescence in U87MG cancer cells in culture</title><author>Horilova, Julia ; Cunderlikova, Beata ; Marcek Chorvatova, Alzbeta</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c493t-55eebbf6389fd2d14e0827840794562ffa3b118768b8aafd9c9f031f9111a8f33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>2,4-Dinitrophenol - chemistry</topic><topic>Cancer</topic><topic>Cell Line, Tumor</topic><topic>Cell Nucleolus - metabolism</topic><topic>Cell Nucleus - metabolism</topic><topic>Confocal</topic><topic>Culture</topic><topic>Deoxyglucose - chemistry</topic><topic>Disease Progression</topic><topic>Early Detection of Cancer - methods</topic><topic>Flavins - chemistry</topic><topic>Fluorescence</topic><topic>Fluorescent Dyes - chemistry</topic><topic>Glycolysis</topic><topic>Humans</topic><topic>Imaging</topic><topic>Microscopy - methods</topic><topic>Microscopy, Confocal</topic><topic>Mitochondria - metabolism</topic><topic>Monitoring</topic><topic>Nuclei</topic><topic>Optics and Photonics - methods</topic><topic>Oxidative Phosphorylation</topic><topic>Oxygen - chemistry</topic><topic>Phosphorylation</topic><topic>Spectra</topic><topic>Spectrometry, Fluorescence - instrumentation</topic><topic>Spectrometry, Fluorescence - methods</topic><topic>Spectrophotometry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Horilova, Julia</creatorcontrib><creatorcontrib>Cunderlikova, Beata</creatorcontrib><creatorcontrib>Marcek Chorvatova, Alzbeta</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Electronics & Communications Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Technology Research Database</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><jtitle>Journal of biomedical optics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Horilova, Julia</au><au>Cunderlikova, Beata</au><au>Marcek Chorvatova, Alzbeta</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Time- and spectrally resolved characteristics of flavin fluorescence in U87MG cancer cells in culture</atitle><jtitle>Journal of biomedical optics</jtitle><addtitle>J. Biomed. Opt</addtitle><date>2015-05-01</date><risdate>2015</risdate><volume>20</volume><issue>5</issue><spage>051017</spage><epage>051017</epage><pages>051017-051017</pages><issn>1083-3668</issn><eissn>1560-2281</eissn><abstract>Early detection of cancer is crucial for the successful diagnostics of its presence and its subsequent treatment. To improve cancer detection, we tested the progressive multimodal optical imaging of U87MG cells in culture. A combination of steady-state spectroscopic methods with the time-resolved approach provides a new insight into the native metabolism when focused on endogenous tissue fluorescence. In this contribution, we evaluated the metabolic state of living U87MG cancer cells in culture by means of endogenous flavin fluorescence. Confocal microscopy and time-resolved fluorescence imaging were employed to gather spectrally and time-resolved images of the flavin fluorescence. We observed that flavin fluorescence in U87MG cells was predominantly localized outside the cell nucleus in mitochondria, while exhibiting a spectral maximum under 500 nm and fluorescence lifetimes under 1.4 ns, suggesting the presence of bound flavins. In some cells, flavin fluorescence was also detected inside the cell nuclei in the nucleoli, exhibiting longer fluorescence lifetimes and a red-shifted spectral maximum, pointing to the presence of free flavin. Extra-nuclear flavin fluorescence was diminished by 2-deoxyglucose, but failed to increase with 2,4-dinitrophenol, the uncoupler of oxidative phosphorylation, indicating that the cells use glycolysis, rather than oxidative phosphorylation for functioning. These gathered data are the first step toward monitoring the metabolic state of U87MG cancer cells.</abstract><cop>United States</cop><pub>Society of Photo-Optical Instrumentation Engineers</pub><pmid>25521208</pmid><doi>10.1117/1.JBO.20.5.051017</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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subjects | 2,4-Dinitrophenol - chemistry Cancer Cell Line, Tumor Cell Nucleolus - metabolism Cell Nucleus - metabolism Confocal Culture Deoxyglucose - chemistry Disease Progression Early Detection of Cancer - methods Flavins - chemistry Fluorescence Fluorescent Dyes - chemistry Glycolysis Humans Imaging Microscopy - methods Microscopy, Confocal Mitochondria - metabolism Monitoring Nuclei Optics and Photonics - methods Oxidative Phosphorylation Oxygen - chemistry Phosphorylation Spectra Spectrometry, Fluorescence - instrumentation Spectrometry, Fluorescence - methods Spectrophotometry |
title | Time- and spectrally resolved characteristics of flavin fluorescence in U87MG cancer cells in culture |
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