Linear Combination Properties of the Phasor Space in Fluorescence Imaging
Resumen:
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.
| 2022 | |
|
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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| Inglés | |
| Universidad de la República | |
| COLIBRI | |
| https://hdl.handle.net/20.500.12008/55177 | |
| Acceso abierto | |
| Licencia Creative Commons Atribución (CC - By 4.0) |
| _version_ | 1877555255850303488 |
|---|---|
| author | Torrado, Belén |
| author2 | Malacrida, Leonel Ranjit, Suman |
| author2_role | author author |
| author_facet | Torrado, Belén Malacrida, Leonel Ranjit, Suman |
| author_role | author |
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| collection | COLIBRI |
| 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.creator.none.fl_str_mv | Torrado, Belén Malacrida, Leonel Ranjit, Suman |
| 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.date.issued.none.fl_str_mv | 2022 |
| dc.description.abstract.none.fl_txt_mv | 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. |
| dc.format.extent.es.fl_str_mv | 27 p. |
| dc.format.mimetype.es.fl_str_mv | application/pdf |
| 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.doi.none.fl_str_mv | 10.3390/s22030999. |
| dc.identifier.eissn.none.fl_str_mv | 1424-8220 |
| dc.identifier.uri.none.fl_str_mv | https://hdl.handle.net/20.500.12008/55177 |
| dc.language.iso.none.fl_str_mv | en eng |
| dc.publisher.es.fl_str_mv | MDPI |
| dc.relation.none.fl_str_mv | Sensors. 2022;22(3) |
| dc.rights.license.none.fl_str_mv | Licencia Creative Commons Atribución (CC - By 4.0) |
| dc.rights.none.fl_str_mv | info:eu-repo/semantics/openAccess |
| dc.source.none.fl_str_mv | reponame:COLIBRI instname:Universidad de la República instacron:Universidad de la República |
| 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 |
| dc.subject.other.es.fl_str_mv | COLORANTES MICROSCOPÍA FLUORESCENTE IMAGEN ÓPTICA |
| dc.title.none.fl_str_mv | Linear Combination Properties of the Phasor Space in Fluorescence Imaging |
| 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 |
| 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. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | COLIBRI_6a18df3bfe7fe8601330c623a8912ce4 |
| 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 |
| instacron_str | Universidad de la República |
| institution | Universidad de la República |
| instname_str | Universidad de la República |
| language | eng |
| language_invalid_str_mv | en |
| network_acronym_str | COLIBRI |
| network_name_str | COLIBRI |
| oai_identifier_str | oai:colibri.udelar.edu.uy:20.500.12008/55177 |
| publishDate | 2022 |
| reponame_str | COLIBRI |
| repository.mail.fl_str_mv | karina.camps@seciu.edu.uy |
| repository.name.fl_str_mv | COLIBRI - Universidad de la República |
| repository_id_str | 4771 |
| rights_invalid_str_mv | Licencia Creative Commons Atribución (CC - By 4.0) |
| 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. Nº 16 de C.D.C. de 07/10/2014)info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución (CC - By 4.0)FLIMFractional intensityHyperspectral imagingLinear combination of phasorModel-freeMultidimensional phasor plotMultiple component analysisPhasorSpectral phasorCOLORANTESMICROSCOPÍA FLUORESCENTEIMAGEN ÓPTICALinear Combination Properties of the Phasor Space in Fluorescence ImagingArtículoinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionreponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaTorrado, BelénMalacrida, LeonelRanjit, SumanLICENSElicense.txtlicense.txttext/plain; charset=utf-84267http://localhost:8080/xmlui/bitstream/20.500.12008/55177/5/license.txt6429389a7df7277b72b7924fdc7d47a9MD55CC-LICENSElicense_urllicense_urltext/plain; charset=utf-844http://localhost:8080/xmlui/bitstream/20.500.12008/55177/2/license_urla0ebbeafb9d2ec7cbb19d7137ebc392cMD52license_textlicense_texttext/html; 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- Universidad de la Repúblicafalse |
| 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 |
| status_str | publishedVersion |
| title | 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_short | Linear Combination Properties of the Phasor Space in Fluorescence Imaging |
| title_sort | Linear Combination Properties of the Phasor Space in Fluorescence Imaging |
| 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 |
| url | https://hdl.handle.net/20.500.12008/55177 |