Linear Combination Properties of the Phasor Space in Fluorescence Imaging

The phasor approach to fluorescence lifetime imaging, and more recently hyperspectral fluorescence imaging, has increased the use of these techniques, and improved the ease and intuitiveness of the data analysis. The fit-free nature of the phasor plots increases the speed of the analysis and reduces...

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Detalhes bibliográficos
Autores: Torrado, Belén, Malacrida, Leonel, Ranjit, Suman
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2022
País:Uruguay
Recursos:Universidad de la República
Repositório:COLIBRI
Idioma:inglês
OAI Identifier:oai:colibri.udelar.edu.uy:20.500.12008/55177
Acesso em linha:https://hdl.handle.net/20.500.12008/55177
Access Level:Acceso aberto
Palavra-chave:FLIM
Fractional intensity
Hyperspectral imaging
Linear combination of phasor
Model-free
Multidimensional phasor plot
Multiple component analysis
Phasor
Spectral phasor
COLORANTES
MICROSCOPÍA FLUORESCENTE
IMAGEN ÓPTICA
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dc.title.none.fl_str_mv Linear Combination Properties of the Phasor Space in Fluorescence Imaging
title Linear Combination Properties of the Phasor Space in Fluorescence Imaging
spellingShingle Linear Combination Properties of the Phasor Space in Fluorescence Imaging
Torrado, Belén
FLIM
Fractional intensity
Hyperspectral imaging
Linear combination of phasor
Model-free
Multidimensional phasor plot
Multiple component analysis
Phasor
Spectral phasor
COLORANTES
MICROSCOPÍA FLUORESCENTE
IMAGEN ÓPTICA
title_short Linear Combination Properties of the Phasor Space in Fluorescence Imaging
title_full Linear Combination Properties of the Phasor Space in Fluorescence Imaging
title_fullStr Linear Combination Properties of the Phasor Space in Fluorescence Imaging
title_full_unstemmed Linear Combination Properties of the Phasor Space in Fluorescence Imaging
title_sort Linear Combination Properties of the Phasor Space in Fluorescence Imaging
dc.creator.none.fl_str_mv Torrado, Belén
Malacrida, Leonel
Ranjit, Suman
author Torrado, Belén
author_facet Torrado, Belén
Malacrida, Leonel
Ranjit, Suman
author_role author
author2 Malacrida, Leonel
Ranjit, Suman
author2_role author
author
dc.contributor.filiacion.none.fl_str_mv Torrado Belén, University of California (E.E.U.U.). Department of Biomedical Engineering. Laboratory for Fluorescence Dynamics
Malacrida Leonel, Universidad de la República (Uruguay). Facultad de Medicina. Departamento de Fisiopatología; Institut Pasteur de Montevideo (Uruguay)
Ranjit Suman, Georgetown University (E.E.U.U.). Department of Biochemistry and Molecular & Cellular Biology, and Microscopy & Imaging Shared Resources
dc.subject.es.fl_str_mv FLIM
Fractional intensity
Hyperspectral imaging
Linear combination of phasor
Model-free
Multidimensional phasor plot
Multiple component analysis
Phasor
Spectral phasor
topic FLIM
Fractional intensity
Hyperspectral imaging
Linear combination of phasor
Model-free
Multidimensional phasor plot
Multiple component analysis
Phasor
Spectral phasor
COLORANTES
MICROSCOPÍA FLUORESCENTE
IMAGEN ÓPTICA
dc.subject.other.es.fl_str_mv COLORANTES
MICROSCOPÍA FLUORESCENTE
IMAGEN ÓPTICA
description The phasor approach to fluorescence lifetime imaging, and more recently hyperspectral fluorescence imaging, has increased the use of these techniques, and improved the ease and intuitiveness of the data analysis. The fit-free nature of the phasor plots increases the speed of the analysis and reduces the dimensionality, optimization of data handling and storage. The reciprocity principle between the real and imaginary space-where the phasor and the pixel that the phasor originated from are linked and can be converted from one another-has helped the expansion of this method. The phasor coordinates calculated from a pixel, where multiple fluorescent species are present, depends on the phasor positions of those components. The relative positions are governed by the linear combination properties of the phasor space. According to this principle, the phasor position of a pixel with multiple components lies inside the polygon whose vertices are occupied by the phasor positions of these individual components and the distance between the image phasor to any of the vertices is inversely proportional to the fractional intensity contribution of that component to the total fluorescence from that image pixel. The higher the fractional intensity contribution of a vertex, the closer is the resultant phasor. The linear additivity in the phasor space can be exploited to obtain the fractional intensity contribution from multiple species and quantify their contribution. This review details the various mathematical models that can be used to obtain two/three/four components from phasor space with known phasor signatures and then how to obtain both the fractional intensities and phasor positions without any prior knowledge of either, assuming they are mono-exponential in nature. We note that other than for blind components, there are no restrictions on the type of the decay or their phasor positions for linear combinations to be valid-and they are applicable to complicated fluorescence lifetime decays from components with intensity decays described by multi-exponentials.
publishDate 2022
dc.date.issued.none.fl_str_mv 2022
dc.date.accessioned.none.fl_str_mv 2026-05-25T18:37:18Z
dc.date.available.none.fl_str_mv 2026-05-25T18:37:18Z
dc.type.es.fl_str_mv Artículo
dc.type.none.fl_str_mv info:eu-repo/semantics/article
dc.type.version.none.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.citation.es.fl_str_mv Torrado B, Malacrida L y Ranjit S. Linear Combination Properties of the Phasor Space in Fluorescence Imaging. Sensors [en línea]. 2022;22(3). 27 p.
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/20.500.12008/55177
dc.identifier.doi.none.fl_str_mv 10.3390/s22030999.
dc.identifier.eissn.none.fl_str_mv 1424-8220
identifier_str_mv Torrado B, Malacrida L y Ranjit S. Linear Combination Properties of the Phasor Space in Fluorescence Imaging. Sensors [en línea]. 2022;22(3). 27 p.
10.3390/s22030999.
1424-8220
url https://hdl.handle.net/20.500.12008/55177
dc.language.iso.none.fl_str_mv en
eng
language_invalid_str_mv en
language eng
dc.relation.none.fl_str_mv Sensors. 2022;22(3)
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
dc.rights.license.none.fl_str_mv Licencia Creative Commons Atribución (CC - By 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv Licencia Creative Commons Atribución (CC - By 4.0)
dc.format.extent.es.fl_str_mv 27 p.
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dc.publisher.es.fl_str_mv MDPI
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instacron_str Universidad de la República
institution Universidad de la República
reponame_str COLIBRI
collection COLIBRI
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spelling Torrado Belén, University of California (E.E.U.U.). Department of Biomedical Engineering. Laboratory for Fluorescence DynamicsMalacrida Leonel, Universidad de la República (Uruguay). Facultad de Medicina. Departamento de Fisiopatología; Institut Pasteur de Montevideo (Uruguay)Ranjit Suman, Georgetown University (E.E.U.U.). Department of Biochemistry and Molecular & Cellular Biology, and Microscopy & Imaging Shared Resources2026-05-25T18:37:18Z2026-05-25T18:37:18Z2022Torrado B, Malacrida L y Ranjit S. Linear Combination Properties of the Phasor Space in Fluorescence Imaging. Sensors [en línea]. 2022;22(3). 27 p.https://hdl.handle.net/20.500.12008/5517710.3390/s22030999.1424-8220The phasor approach to fluorescence lifetime imaging, and more recently hyperspectral fluorescence imaging, has increased the use of these techniques, and improved the ease and intuitiveness of the data analysis. The fit-free nature of the phasor plots increases the speed of the analysis and reduces the dimensionality, optimization of data handling and storage. The reciprocity principle between the real and imaginary space-where the phasor and the pixel that the phasor originated from are linked and can be converted from one another-has helped the expansion of this method. The phasor coordinates calculated from a pixel, where multiple fluorescent species are present, depends on the phasor positions of those components. The relative positions are governed by the linear combination properties of the phasor space. According to this principle, the phasor position of a pixel with multiple components lies inside the polygon whose vertices are occupied by the phasor positions of these individual components and the distance between the image phasor to any of the vertices is inversely proportional to the fractional intensity contribution of that component to the total fluorescence from that image pixel. The higher the fractional intensity contribution of a vertex, the closer is the resultant phasor. The linear additivity in the phasor space can be exploited to obtain the fractional intensity contribution from multiple species and quantify their contribution. This review details the various mathematical models that can be used to obtain two/three/four components from phasor space with known phasor signatures and then how to obtain both the fractional intensities and phasor positions without any prior knowledge of either, assuming they are mono-exponential in nature. We note that other than for blind components, there are no restrictions on the type of the decay or their phasor positions for linear combinations to be valid-and they are applicable to complicated fluorescence lifetime decays from components with intensity decays described by multi-exponentials.Submitted by Almiñana María Cecilia (marialminana@gmail.com) on 2026-05-25T17:42:15Z No. of bitstreams: 2 license_rdf: 25630 bytes, checksum: e7132498e7c1fe99f7096667baa99b25 (MD5) Linear Combination Properties of the Phasor Space.pdf: 39400773 bytes, checksum: edf0f6749b7a7412f5a26cf246dfd88f (MD5)Approved for entry into archive by Almiñana María Cecilia (marialminana@gmail.com) on 2026-05-25T17:57:08Z (GMT) No. of bitstreams: 2 license_rdf: 25630 bytes, checksum: e7132498e7c1fe99f7096667baa99b25 (MD5) Linear Combination Properties of the Phasor Space.pdf: 39400773 bytes, checksum: edf0f6749b7a7412f5a26cf246dfd88f (MD5)Made available in DSpace by Camps Karina (karina.camps@seciu.edu.uy) on 2026-05-25T18:37:18Z (GMT). No. of bitstreams: 2 license_rdf: 25630 bytes, checksum: e7132498e7c1fe99f7096667baa99b25 (MD5) Linear Combination Properties of the Phasor Space.pdf: 39400773 bytes, checksum: edf0f6749b7a7412f5a26cf246dfd88f (MD5) Previous issue date: 202227 p.application/pdfenengMDPISensors. 2022;22(3)Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad de la República.(Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. 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públicahttps://udelar.edu.uy/https://www.colibri.udelar.edu.uy/oai/requestkarina.camps@seciu.edu.uyUruguayopendoar:47712026-05-25T18:37:18COLIBRI - Universidad de la Repúblicafalse
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