Multiplexed Analysis of Proteins in Tissue Using Multispectral Fluorescence Imaging
We present a new application of multispectral analysis for subcellular measurement of multiple proteins in formalin-fixed paraffin embedded tissue and cells. Typically, the targets of interest are present in the same or spatially overlapping cellular compartments. Such co-localization can complicate...
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| Veröffentlicht in: | IEEE transactions on medical imaging 2010-08, Vol.29 (8), p.1457-1462 |
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| container_title | IEEE transactions on medical imaging |
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| creator | Barash, E Dinn, S Sevinsky, C Ginty, F |
| description | We present a new application of multispectral analysis for subcellular measurement of multiple proteins in formalin-fixed paraffin embedded tissue and cells. Typically, the targets of interest are present in the same or spatially overlapping cellular compartments. Such co-localization can complicate analysis and interpretation of the images obtained using traditional fluorescence, especially when spectrally overlapping labels are present. The spectral properties of currently available fluorescent dyes set an upper limit to the number of molecules that can be detected simultaneously with traditional fluorescence. By exciting a set of fluorophores at the same wavelength and unmixing their emission signals from background autofluorescence, we were able to image three targets in a single channel. This parallel imaging approach provides significant advantages for multiplexed analysis of tissues and cells. |
| doi_str_mv | 10.1109/TMI.2010.2045005 |
| format | Article |
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Typically, the targets of interest are present in the same or spatially overlapping cellular compartments. Such co-localization can complicate analysis and interpretation of the images obtained using traditional fluorescence, especially when spectrally overlapping labels are present. The spectral properties of currently available fluorescent dyes set an upper limit to the number of molecules that can be detected simultaneously with traditional fluorescence. By exciting a set of fluorophores at the same wavelength and unmixing their emission signals from background autofluorescence, we were able to image three targets in a single channel. This parallel imaging approach provides significant advantages for multiplexed analysis of tissues and cells.</description><identifier>ISSN: 0278-0062</identifier><identifier>EISSN: 1558-254X</identifier><identifier>DOI: 10.1109/TMI.2010.2045005</identifier><identifier>PMID: 20304722</identifier><identifier>CODEN: ITMID4</identifier><language>eng</language><publisher>United States: IEEE</publisher><subject>Algorithms ; Animals ; Assembly ; Band pass filters ; Biomarkers ; Biomarkers - analysis ; Breast - chemistry ; Breast Neoplasms - metabolism ; Carbocyanines - chemistry ; Channels ; Compartments ; Cyclic AMP Response Element-Binding Protein - analysis ; Female ; Fluorescence ; Humans ; Image analysis ; Image Processing, Computer-Assisted - methods ; Imaging ; Immunofluorescence ; Lung - chemistry ; Male ; Mice ; Microscopy ; Microscopy, Fluorescence - methods ; Microscopy, Fluorescence, Multiphoton - methods ; Molecular imaging ; Multiplexing ; multispectral imaging ; Optical imaging ; Pharmaceutical technology ; Prostate - chemistry ; protein multiplexes and autofluorescence ; Proteins ; Proteins - analysis ; Signal Processing, Computer-Assisted ; Spectra ; Tissue Array Analysis - methods ; Wavelengths</subject><ispartof>IEEE transactions on medical imaging, 2010-08, Vol.29 (8), p.1457-1462</ispartof><rights>Copyright The Institute of Electrical and Electronics Engineers, Inc. (IEEE) Aug 2010</rights><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c411t-229a017cd71ed427b97590dff716141a3c9a55f61c346e22d825ab5b2898f24a3</citedby><cites>FETCH-LOGICAL-c411t-229a017cd71ed427b97590dff716141a3c9a55f61c346e22d825ab5b2898f24a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://ieeexplore.ieee.org/document/5433073$$EHTML$$P50$$Gieee$$H</linktohtml><link.rule.ids>315,781,785,797,27928,27929,54762</link.rule.ids><linktorsrc>$$Uhttps://ieeexplore.ieee.org/document/5433073$$EView_record_in_IEEE$$FView_record_in_$$GIEEE</linktorsrc><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20304722$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Barash, E</creatorcontrib><creatorcontrib>Dinn, S</creatorcontrib><creatorcontrib>Sevinsky, C</creatorcontrib><creatorcontrib>Ginty, F</creatorcontrib><title>Multiplexed Analysis of Proteins in Tissue Using Multispectral Fluorescence Imaging</title><title>IEEE transactions on medical imaging</title><addtitle>TMI</addtitle><addtitle>IEEE Trans Med Imaging</addtitle><description>We present a new application of multispectral analysis for subcellular measurement of multiple proteins in formalin-fixed paraffin embedded tissue and cells. Typically, the targets of interest are present in the same or spatially overlapping cellular compartments. Such co-localization can complicate analysis and interpretation of the images obtained using traditional fluorescence, especially when spectrally overlapping labels are present. The spectral properties of currently available fluorescent dyes set an upper limit to the number of molecules that can be detected simultaneously with traditional fluorescence. By exciting a set of fluorophores at the same wavelength and unmixing their emission signals from background autofluorescence, we were able to image three targets in a single channel. This parallel imaging approach provides significant advantages for multiplexed analysis of tissues and cells.</description><subject>Algorithms</subject><subject>Animals</subject><subject>Assembly</subject><subject>Band pass filters</subject><subject>Biomarkers</subject><subject>Biomarkers - analysis</subject><subject>Breast - chemistry</subject><subject>Breast Neoplasms - metabolism</subject><subject>Carbocyanines - chemistry</subject><subject>Channels</subject><subject>Compartments</subject><subject>Cyclic AMP Response Element-Binding Protein - analysis</subject><subject>Female</subject><subject>Fluorescence</subject><subject>Humans</subject><subject>Image analysis</subject><subject>Image Processing, Computer-Assisted - methods</subject><subject>Imaging</subject><subject>Immunofluorescence</subject><subject>Lung - chemistry</subject><subject>Male</subject><subject>Mice</subject><subject>Microscopy</subject><subject>Microscopy, Fluorescence - methods</subject><subject>Microscopy, Fluorescence, Multiphoton - methods</subject><subject>Molecular imaging</subject><subject>Multiplexing</subject><subject>multispectral imaging</subject><subject>Optical imaging</subject><subject>Pharmaceutical technology</subject><subject>Prostate - chemistry</subject><subject>protein multiplexes and autofluorescence</subject><subject>Proteins</subject><subject>Proteins - analysis</subject><subject>Signal Processing, Computer-Assisted</subject><subject>Spectra</subject><subject>Tissue Array Analysis - methods</subject><subject>Wavelengths</subject><issn>0278-0062</issn><issn>1558-254X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>RIE</sourceid><sourceid>EIF</sourceid><recordid>eNqF0cFrFDEUBvAgil2rd0GQgAe9TH15SSaTYym2XWip4Ba8DdmZNyVldmZNZqD9733trj30UCEQQn7vI-QT4qOCI6XAf19dLo8Q-IRgLIB9JRbK2qpAa36_FgtAVxUAJR6IdznfAihW_q04QNBgHOJC_Lqc-ylue7qjVh4Pob_PMcuxkz_TOFEcsoyDXMWcZ5LXOQ438nEgb6mZUujlaT-PiXJDQ0NyuQk3TN6LN13oM33Y74fi-vTH6uS8uLg6W54cXxSNUWoqEH0A5ZrWKWoNurV31kPbdU6VyqigGx-s7UrVaFMSYluhDWu7xspXHZqgD8XXXe42jX9mylO9ifySvg8DjXOuOU5zkDP_l6by2vNi-e1FqUqnEK33iumXZ_R2nBN_ISv-eecqB5YV7FSTxpwTdfU2xU1I94zqhxJrLrF-KLHel8gjn_fB83pD7dPAv9YYfNqBSERP19ZoDU7rv2TkniU</recordid><startdate>201008</startdate><enddate>201008</enddate><creator>Barash, E</creator><creator>Dinn, S</creator><creator>Sevinsky, C</creator><creator>Ginty, F</creator><general>IEEE</general><general>The Institute of Electrical and Electronics Engineers, Inc. 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analysis</topic><topic>Breast - chemistry</topic><topic>Breast Neoplasms - metabolism</topic><topic>Carbocyanines - chemistry</topic><topic>Channels</topic><topic>Compartments</topic><topic>Cyclic AMP Response Element-Binding Protein - analysis</topic><topic>Female</topic><topic>Fluorescence</topic><topic>Humans</topic><topic>Image analysis</topic><topic>Image Processing, Computer-Assisted - methods</topic><topic>Imaging</topic><topic>Immunofluorescence</topic><topic>Lung - chemistry</topic><topic>Male</topic><topic>Mice</topic><topic>Microscopy</topic><topic>Microscopy, Fluorescence - methods</topic><topic>Microscopy, Fluorescence, Multiphoton - methods</topic><topic>Molecular imaging</topic><topic>Multiplexing</topic><topic>multispectral imaging</topic><topic>Optical imaging</topic><topic>Pharmaceutical technology</topic><topic>Prostate - chemistry</topic><topic>protein multiplexes and autofluorescence</topic><topic>Proteins</topic><topic>Proteins - analysis</topic><topic>Signal Processing, Computer-Assisted</topic><topic>Spectra</topic><topic>Tissue Array Analysis - 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Academic</collection><jtitle>IEEE transactions on medical imaging</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext_linktorsrc</fulltext></delivery><addata><au>Barash, E</au><au>Dinn, S</au><au>Sevinsky, C</au><au>Ginty, F</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Multiplexed Analysis of Proteins in Tissue Using Multispectral Fluorescence Imaging</atitle><jtitle>IEEE transactions on medical imaging</jtitle><stitle>TMI</stitle><addtitle>IEEE Trans Med Imaging</addtitle><date>2010-08</date><risdate>2010</risdate><volume>29</volume><issue>8</issue><spage>1457</spage><epage>1462</epage><pages>1457-1462</pages><issn>0278-0062</issn><eissn>1558-254X</eissn><coden>ITMID4</coden><abstract>We present a new application of multispectral analysis for subcellular measurement of multiple proteins in formalin-fixed paraffin embedded tissue and cells. Typically, the targets of interest are present in the same or spatially overlapping cellular compartments. Such co-localization can complicate analysis and interpretation of the images obtained using traditional fluorescence, especially when spectrally overlapping labels are present. The spectral properties of currently available fluorescent dyes set an upper limit to the number of molecules that can be detected simultaneously with traditional fluorescence. By exciting a set of fluorophores at the same wavelength and unmixing their emission signals from background autofluorescence, we were able to image three targets in a single channel. This parallel imaging approach provides significant advantages for multiplexed analysis of tissues and cells.</abstract><cop>United States</cop><pub>IEEE</pub><pmid>20304722</pmid><doi>10.1109/TMI.2010.2045005</doi><tpages>6</tpages></addata></record> |
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| subjects | Algorithms Animals Assembly Band pass filters Biomarkers Biomarkers - analysis Breast - chemistry Breast Neoplasms - metabolism Carbocyanines - chemistry Channels Compartments Cyclic AMP Response Element-Binding Protein - analysis Female Fluorescence Humans Image analysis Image Processing, Computer-Assisted - methods Imaging Immunofluorescence Lung - chemistry Male Mice Microscopy Microscopy, Fluorescence - methods Microscopy, Fluorescence, Multiphoton - methods Molecular imaging Multiplexing multispectral imaging Optical imaging Pharmaceutical technology Prostate - chemistry protein multiplexes and autofluorescence Proteins Proteins - analysis Signal Processing, Computer-Assisted Spectra Tissue Array Analysis - methods Wavelengths |
| title | Multiplexed Analysis of Proteins in Tissue Using Multispectral Fluorescence Imaging |
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