Phasor approach to fluorescence lifetime microscopy distinguishes different metabolic states of germ cells in a live tissue
We describe a label-free imaging method to monitor stem-cell metabolism that discriminates different states of stem cells as they differentiate in living tissues. In this method we use intrinsic fluorescence biomarkers and the phasor approach to fluorescence lifetime imaging microscopy in conjunctio...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2011-08, Vol.108 (33), p.13582-13587 |
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| container_issue | 33 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Stringari, Chiara Cinquin, Amanda Cinquin, Olivier Digman, Michelle A Donovan, Peter J Gratton, Enrico |
| description | We describe a label-free imaging method to monitor stem-cell metabolism that discriminates different states of stem cells as they differentiate in living tissues. In this method we use intrinsic fluorescence biomarkers and the phasor approach to fluorescence lifetime imaging microscopy in conjunction with image segmentation, which we use to introduce the concept of the cell phasor. In live tissues we are able to identify intrinsic fluorophores, such as collagen, retinol, retinoic acid, porphyrin, flavins, and free and bound NADH. We have exploited the cell phasor approach to detect a trend in metabolite concentrations along the main axis of the Caenorhabditis elegans germ line. This trend is consistent with known changes in metabolic states during differentiation. The cell phasor approach to lifetime imaging provides a label-free, fit-free, and sensitive method to identify different metabolic states of cells during differentiation, to sense small changes in the redox state of cells, and may identify symmetric and asymmetric divisions and predict cell fate. Our method is a promising noninvasive optical tool for monitoring metabolic pathways during differentiation or disease progression, and for cell sorting in unlabeled tissues. |
| doi_str_mv | 10.1073/pnas.1108161108 |
| format | Article |
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In this method we use intrinsic fluorescence biomarkers and the phasor approach to fluorescence lifetime imaging microscopy in conjunction with image segmentation, which we use to introduce the concept of the cell phasor. In live tissues we are able to identify intrinsic fluorophores, such as collagen, retinol, retinoic acid, porphyrin, flavins, and free and bound NADH. We have exploited the cell phasor approach to detect a trend in metabolite concentrations along the main axis of the Caenorhabditis elegans germ line. This trend is consistent with known changes in metabolic states during differentiation. The cell phasor approach to lifetime imaging provides a label-free, fit-free, and sensitive method to identify different metabolic states of cells during differentiation, to sense small changes in the redox state of cells, and may identify symmetric and asymmetric divisions and predict cell fate. Our method is a promising noninvasive optical tool for monitoring metabolic pathways during differentiation or disease progression, and for cell sorting in unlabeled tissues.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.1108161108</identifier><identifier>PMID: 21808026</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>Animals ; biochemical pathways ; Biological Sciences ; Biomarkers ; Caenorhabditis elegans ; Caenorhabditis elegans - cytology ; Cell Differentiation ; Cellular differentiation ; Cellular metabolism ; collagen ; Collagens ; disease course ; flavins ; Fluorescence ; Fluorescent Dyes ; Germ cells ; Germ Cells - cytology ; Germ Cells - metabolism ; Half-Life ; image analysis ; Imaging ; Metabolism ; metabolites ; Metabolomics - methods ; Methods ; Microscopy ; Microscopy, Fluorescence - methods ; monitoring ; NAD (coenzyme) ; Nematodes ; Nonnative species ; Pixels ; Pluripotent stem cells ; retinoic acid ; Stem cells ; vitamin A</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2011-08, Vol.108 (33), p.13582-13587</ispartof><rights>copyright © 1993–2008 National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences Aug 16, 2011</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c556t-3847e1fcd5fdf7a2114523c0228c1003fd446c9e3fc587ff66cd90a09683a20f3</citedby><cites>FETCH-LOGICAL-c556t-3847e1fcd5fdf7a2114523c0228c1003fd446c9e3fc587ff66cd90a09683a20f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/108/33.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/27979241$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/27979241$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,727,780,784,803,885,27923,27924,53790,53792,58016,58249</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21808026$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Stringari, Chiara</creatorcontrib><creatorcontrib>Cinquin, Amanda</creatorcontrib><creatorcontrib>Cinquin, Olivier</creatorcontrib><creatorcontrib>Digman, Michelle A</creatorcontrib><creatorcontrib>Donovan, Peter J</creatorcontrib><creatorcontrib>Gratton, Enrico</creatorcontrib><title>Phasor approach to fluorescence lifetime microscopy distinguishes different metabolic states of germ cells in a live tissue</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>We describe a label-free imaging method to monitor stem-cell metabolism that discriminates different states of stem cells as they differentiate in living tissues. 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Our method is a promising noninvasive optical tool for monitoring metabolic pathways during differentiation or disease progression, and for cell sorting in unlabeled tissues.</description><subject>Animals</subject><subject>biochemical pathways</subject><subject>Biological Sciences</subject><subject>Biomarkers</subject><subject>Caenorhabditis elegans</subject><subject>Caenorhabditis elegans - cytology</subject><subject>Cell Differentiation</subject><subject>Cellular differentiation</subject><subject>Cellular metabolism</subject><subject>collagen</subject><subject>Collagens</subject><subject>disease course</subject><subject>flavins</subject><subject>Fluorescence</subject><subject>Fluorescent Dyes</subject><subject>Germ cells</subject><subject>Germ Cells - cytology</subject><subject>Germ Cells - metabolism</subject><subject>Half-Life</subject><subject>image analysis</subject><subject>Imaging</subject><subject>Metabolism</subject><subject>metabolites</subject><subject>Metabolomics - methods</subject><subject>Methods</subject><subject>Microscopy</subject><subject>Microscopy, Fluorescence - methods</subject><subject>monitoring</subject><subject>NAD (coenzyme)</subject><subject>Nematodes</subject><subject>Nonnative species</subject><subject>Pixels</subject><subject>Pluripotent stem cells</subject><subject>retinoic acid</subject><subject>Stem cells</subject><subject>vitamin A</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkcFvFCEYxYnR2LV69qQSL562_YCBgYuJaWw1aaKJ9kwoA7tsZoYRmCaN_7yMu3bVC4S8Hy_f-x5CLwmcEWjZ-TSafEYISCKW8xFaEVBkLRoFj9EKgLZr2dDmBD3LeQcAikt4ik4okSCBihX6-XVrckzYTFOKxm5xidj3c0wuWzdah_vgXQmDw0OwKWYbp3vchVzCuJlD3rpcX9675MaCB1fMbeyDxbmYUqXo8calAVvX9xmHEZvqd-dwCTnP7jl64k2f3YvDfYpuLj9-v_i0vv5y9fniw_Xaci7KmsmmdcTbjvvOt4YS0nDKLFAqLQFgvmsaYZVj3nLZei-E7RQYUEIyQ8GzU_R-7zvNt4Praq6STK-nFAaT7nU0Qf-rjGGrN_FOM8Il4aIavDsYpPhjdrnoIeQlkxldnLOWdcdC8RYq-fY_chfnNNZ0vyFVZ1IVOt9Dy0Jzcv5hFAJ6qVUvtepjrfXH678TPPB_eqwAPgDLz6Od1IxpwrikFXm1R3a5xHS0aFWraEOq_mavexO12aSQ9c03CqQBIAoYI-wX1WO-gw</recordid><startdate>20110816</startdate><enddate>20110816</enddate><creator>Stringari, Chiara</creator><creator>Cinquin, Amanda</creator><creator>Cinquin, Olivier</creator><creator>Digman, Michelle A</creator><creator>Donovan, Peter J</creator><creator>Gratton, Enrico</creator><general>National Academy of Sciences</general><general>National Acad Sciences</general><scope>FBQ</scope><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>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20110816</creationdate><title>Phasor approach to fluorescence lifetime microscopy distinguishes different metabolic states of germ cells in a live tissue</title><author>Stringari, Chiara ; 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In this method we use intrinsic fluorescence biomarkers and the phasor approach to fluorescence lifetime imaging microscopy in conjunction with image segmentation, which we use to introduce the concept of the cell phasor. In live tissues we are able to identify intrinsic fluorophores, such as collagen, retinol, retinoic acid, porphyrin, flavins, and free and bound NADH. We have exploited the cell phasor approach to detect a trend in metabolite concentrations along the main axis of the Caenorhabditis elegans germ line. This trend is consistent with known changes in metabolic states during differentiation. The cell phasor approach to lifetime imaging provides a label-free, fit-free, and sensitive method to identify different metabolic states of cells during differentiation, to sense small changes in the redox state of cells, and may identify symmetric and asymmetric divisions and predict cell fate. Our method is a promising noninvasive optical tool for monitoring metabolic pathways during differentiation or disease progression, and for cell sorting in unlabeled tissues.</abstract><cop>United States</cop><pub>National Academy of Sciences</pub><pmid>21808026</pmid><doi>10.1073/pnas.1108161108</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals biochemical pathways Biological Sciences Biomarkers Caenorhabditis elegans Caenorhabditis elegans - cytology Cell Differentiation Cellular differentiation Cellular metabolism collagen Collagens disease course flavins Fluorescence Fluorescent Dyes Germ cells Germ Cells - cytology Germ Cells - metabolism Half-Life image analysis Imaging Metabolism metabolites Metabolomics - methods Methods Microscopy Microscopy, Fluorescence - methods monitoring NAD (coenzyme) Nematodes Nonnative species Pixels Pluripotent stem cells retinoic acid Stem cells vitamin A |
| title | Phasor approach to fluorescence lifetime microscopy distinguishes different metabolic states of germ cells in a live tissue |
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