A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging

A coumarin fluorophore and an oxazine photochrome can be integrated within the same molecular skeleton and connected covalently to a secondary antibody. Illumination of the antibody-dyad conjugate at an appropriate activation wavelength opens the oxazine ring reversibly and shifts bathochromically t...

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Detalles Bibliográficos
Autores principales:	Roberti, Maria Julia, Araoz, Beatriz, Bossi, Mariano Luis
Publicado:	2016
Materias:	Biomolecules Chemical activation Chromophores Excited states Fluorescence Ground state Molecules Nitrogen compounds Optical resolving power Fluorescence switching Ground state absorption Molecular skeleton Operating principles Secondary antibodies Selective excitations Single molecule level Super resolution imaging Antibodies
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19327447_v120_n23_p12860_Cusido http://hdl.handle.net/20.500.12110/paper_19327447_v120_n23_p12860_Cusido
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_19327447_v120_n23_p12860_Cusido
record_format	dspace
spelling	paper:paper_19327447_v120_n23_p12860_Cusido2023-06-08T16:31:39Z A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging Roberti, Maria Julia Araoz, Beatriz Bossi, Mariano Luis Biomolecules Chemical activation Chromophores Excited states Fluorescence Ground state Molecules Nitrogen compounds Optical resolving power Fluorescence switching Ground state absorption Molecular skeleton Operating principles Secondary antibodies Selective excitations Single molecule level Super resolution imaging Antibodies A coumarin fluorophore and an oxazine photochrome can be integrated within the same molecular skeleton and connected covalently to a secondary antibody. Illumination of the antibody-dyad conjugate at an appropriate activation wavelength opens the oxazine ring reversibly and shifts bathochromically the ground-state absorption of the coumarin component. Selective excitation of the photochemical product then produces significant fluorescence and allows the detection of activated bioconjugates at the single-molecule level. Such fluorescence activation events can be exploited to resolve temporally individual emitters and reconstruct images of immunolabeled cells with subdiffraction resolution. Relying on a similar conjugation protocol, a model compound, incorporating the same chromophore of the photochemical product, can also be connected covalently to a secondary antibody. Stimulated emission can be exploited to deplete the excited state of the bioconjugated chromophores and switch their fluorescence off. These operating principles for fluorescence switching also permit the imaging of immunolabeled cells with subdiffraction resolution. Thus, these photoswitchable molecules, in combination with the labeling ability of antibodies, can evolve into valuable probes for bioimaging with superresolution. © 2016 American Chemical Society. Fil:Roberti, M.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Araoz, B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Bossi, M.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2016 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19327447_v120_n23_p12860_Cusido http://hdl.handle.net/20.500.12110/paper_19327447_v120_n23_p12860_Cusido
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Biomolecules Chemical activation Chromophores Excited states Fluorescence Ground state Molecules Nitrogen compounds Optical resolving power Fluorescence switching Ground state absorption Molecular skeleton Operating principles Secondary antibodies Selective excitations Single molecule level Super resolution imaging Antibodies
spellingShingle	Biomolecules Chemical activation Chromophores Excited states Fluorescence Ground state Molecules Nitrogen compounds Optical resolving power Fluorescence switching Ground state absorption Molecular skeleton Operating principles Secondary antibodies Selective excitations Single molecule level Super resolution imaging Antibodies Roberti, Maria Julia Araoz, Beatriz Bossi, Mariano Luis A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging
topic_facet	Biomolecules Chemical activation Chromophores Excited states Fluorescence Ground state Molecules Nitrogen compounds Optical resolving power Fluorescence switching Ground state absorption Molecular skeleton Operating principles Secondary antibodies Selective excitations Single molecule level Super resolution imaging Antibodies
description	A coumarin fluorophore and an oxazine photochrome can be integrated within the same molecular skeleton and connected covalently to a secondary antibody. Illumination of the antibody-dyad conjugate at an appropriate activation wavelength opens the oxazine ring reversibly and shifts bathochromically the ground-state absorption of the coumarin component. Selective excitation of the photochemical product then produces significant fluorescence and allows the detection of activated bioconjugates at the single-molecule level. Such fluorescence activation events can be exploited to resolve temporally individual emitters and reconstruct images of immunolabeled cells with subdiffraction resolution. Relying on a similar conjugation protocol, a model compound, incorporating the same chromophore of the photochemical product, can also be connected covalently to a secondary antibody. Stimulated emission can be exploited to deplete the excited state of the bioconjugated chromophores and switch their fluorescence off. These operating principles for fluorescence switching also permit the imaging of immunolabeled cells with subdiffraction resolution. Thus, these photoswitchable molecules, in combination with the labeling ability of antibodies, can evolve into valuable probes for bioimaging with superresolution. © 2016 American Chemical Society.
author	Roberti, Maria Julia Araoz, Beatriz Bossi, Mariano Luis
author_facet	Roberti, Maria Julia Araoz, Beatriz Bossi, Mariano Luis
author_sort	Roberti, Maria Julia
title	A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging
title_short	A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging
title_full	A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging
title_fullStr	A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging
title_full_unstemmed	A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging
title_sort	photochromic bioconjugate with photoactivatable fluorescence for superresolution imaging
publishDate	2016
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19327447_v120_n23_p12860_Cusido http://hdl.handle.net/20.500.12110/paper_19327447_v120_n23_p12860_Cusido
work_keys_str_mv	AT robertimariajulia aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT araozbeatriz aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT bossimarianoluis aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT robertimariajulia photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT araozbeatriz photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT bossimarianoluis photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging
_version_	1768543104805109760

A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging

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