Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications
- Autores
- Belov, Vladimir N.; Mitronova, Gyuzel Yu.; Bossi, Mariano Luis; Boyarskiy, Vadim P.; Hebisch, Elke; Geisler, Claudia; Kolmakov, Kirill; Wurm, Christian A.; Willig, Katrin I.; Hell, Stefan W.
- Año de publicación
- 2014
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- Caged rhodamine dyes (Rhodamines NN) of five basic colors were synthesized and used as “hidden” markers in subdiffractional and conventional light microscopy. These masked fluorophores with a 2-diazo-1-indanone group can be irreversibly photoactivated, either by irradiation with UV- or violet light (one-photon process), or by exposure to intense red light (λ∼750 nm; two-photon mode). All dyes possess a very small 2-diazoketone caging group incorporated into the 2-diazo-1-indanone residue with a quaternary carbon atom (C-3) and a spiro-9H-xanthene fragment. Initially they are non-colored (pale yellow), non-fluorescent, and absorb at λ=330–350 nm (molar extinction coefficient (ε)≈104 M−1 cm−1) with a band edge that extends to about λ=440 nm. The absorption and emission bands of the uncaged derivatives are tunable over a wide range (λ=511–633 and 525–653 nm, respectively). The unmasked dyes are highly colored and fluorescent (ε= 3–8×104 M−1 cm−1 and fluorescence quantum yields (ϕ)=40–85 % in the unbound state and in methanol). By stepwise and orthogonal protection of carboxylic and sulfonic acid groups a highly water-soluble caged red-emitting dye with two sulfonic acid residues was prepared. Rhodamines NN were decorated with amino-reactive N-hydroxysuccinimidyl ester groups, applied in aqueous buffers, easily conjugated with proteins, and readily photoactivated (uncaged) with λ=375–420 nm light or intense red light (λ=775 nm). Protein conjugates with optimal degrees of labeling (3–6) were prepared and uncaged with λ=405 nm light in aqueous buffer solutions (ϕ=20–38 %). The photochemical cleavage of the masking group generates only molecular nitrogen. Some 10–40 % of the non-fluorescent (dark) byproducts are also formed. However, they have low absorbance and do not quench the fluorescence of the uncaged dyes. Photoactivation of the individual molecules of Rhodamines NN (e.g., due to reversible or irreversible transition to a “dark” non-emitting state or photobleaching) provides multicolor images with subdiffractional optical resolution. The applicability of these novel caged fluorophores in super-resolution optical microscopy is exemplified.
Fil: Belov, Vladimir N.. Max Planck Institute for Biophysical Chemistry ; Alemania
Fil: Mitronova, Gyuzel Yu.. Max Planck Institute for Biophysical Chemistry ; Alemania
Fil: Bossi, Mariano Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina
Fil: Boyarskiy, Vadim P.. St. Petersburg State University; Rusia
Fil: Hebisch, Elke. Max Planck Institute for Biophysical Chemistry ; Alemania
Fil: Geisler, Claudia. Max Planck Institute for Biophysical Chemistry ; Alemania
Fil: Kolmakov, Kirill. Max Planck Institute for Biophysical Chemistry ; Alemania
Fil: Wurm, Christian A.. Max Planck Institute for Biophysical Chemistry ; Alemania
Fil: Willig, Katrin I.. Max Planck Institute for Biophysical Chemistry ; Alemania
Fil: Hell, Stefan W.. Max Planck Institute for Biophysical Chemistry ; Alemania - Materia
-
Bioconjugation
Fluorescence
Photolysis
Rhodamines - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/32179
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Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy ApplicationsBelov, Vladimir N.Mitronova, Gyuzel Yu.Bossi, Mariano LuisBoyarskiy, Vadim P.Hebisch, ElkeGeisler, ClaudiaKolmakov, KirillWurm, Christian A.Willig, Katrin I.Hell, Stefan W.BioconjugationFluorescencePhotolysisRhodamineshttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Caged rhodamine dyes (Rhodamines NN) of five basic colors were synthesized and used as “hidden” markers in subdiffractional and conventional light microscopy. These masked fluorophores with a 2-diazo-1-indanone group can be irreversibly photoactivated, either by irradiation with UV- or violet light (one-photon process), or by exposure to intense red light (λ∼750 nm; two-photon mode). All dyes possess a very small 2-diazoketone caging group incorporated into the 2-diazo-1-indanone residue with a quaternary carbon atom (C-3) and a spiro-9H-xanthene fragment. Initially they are non-colored (pale yellow), non-fluorescent, and absorb at λ=330–350 nm (molar extinction coefficient (ε)≈104 M−1 cm−1) with a band edge that extends to about λ=440 nm. The absorption and emission bands of the uncaged derivatives are tunable over a wide range (λ=511–633 and 525–653 nm, respectively). The unmasked dyes are highly colored and fluorescent (ε= 3–8×104 M−1 cm−1 and fluorescence quantum yields (ϕ)=40–85 % in the unbound state and in methanol). By stepwise and orthogonal protection of carboxylic and sulfonic acid groups a highly water-soluble caged red-emitting dye with two sulfonic acid residues was prepared. Rhodamines NN were decorated with amino-reactive N-hydroxysuccinimidyl ester groups, applied in aqueous buffers, easily conjugated with proteins, and readily photoactivated (uncaged) with λ=375–420 nm light or intense red light (λ=775 nm). Protein conjugates with optimal degrees of labeling (3–6) were prepared and uncaged with λ=405 nm light in aqueous buffer solutions (ϕ=20–38 %). The photochemical cleavage of the masking group generates only molecular nitrogen. Some 10–40 % of the non-fluorescent (dark) byproducts are also formed. However, they have low absorbance and do not quench the fluorescence of the uncaged dyes. Photoactivation of the individual molecules of Rhodamines NN (e.g., due to reversible or irreversible transition to a “dark” non-emitting state or photobleaching) provides multicolor images with subdiffractional optical resolution. The applicability of these novel caged fluorophores in super-resolution optical microscopy is exemplified.Fil: Belov, Vladimir N.. Max Planck Institute for Biophysical Chemistry ; AlemaniaFil: Mitronova, Gyuzel Yu.. Max Planck Institute for Biophysical Chemistry ; AlemaniaFil: Bossi, Mariano Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; ArgentinaFil: Boyarskiy, Vadim P.. St. Petersburg State University; RusiaFil: Hebisch, Elke. Max Planck Institute for Biophysical Chemistry ; AlemaniaFil: Geisler, Claudia. Max Planck Institute for Biophysical Chemistry ; AlemaniaFil: Kolmakov, Kirill. Max Planck Institute for Biophysical Chemistry ; AlemaniaFil: Wurm, Christian A.. Max Planck Institute for Biophysical Chemistry ; AlemaniaFil: Willig, Katrin I.. Max Planck Institute for Biophysical Chemistry ; AlemaniaFil: Hell, Stefan W.. Max Planck Institute for Biophysical Chemistry ; AlemaniaWiley VCH Verlag2014-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/32179Belov, Vladimir N.; Hebisch, Elke; Mitronova, Gyuzel Yu.; Bossi, Mariano Luis; Boyarskiy, Vadim P.; Geisler, Claudia; et al.; Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications; Wiley VCH Verlag; Chemistry- A European Journal; 20; 41; 9-2014; 13162-131730947-6539CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1002/chem.201403316info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1002/chem.201403316/abstractinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:59:48Zoai:ri.conicet.gov.ar:11336/32179instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:59:48.614CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications |
title |
Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications |
spellingShingle |
Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications Belov, Vladimir N. Bioconjugation Fluorescence Photolysis Rhodamines |
title_short |
Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications |
title_full |
Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications |
title_fullStr |
Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications |
title_full_unstemmed |
Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications |
title_sort |
Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications |
dc.creator.none.fl_str_mv |
Belov, Vladimir N. Mitronova, Gyuzel Yu. Bossi, Mariano Luis Boyarskiy, Vadim P. Hebisch, Elke Geisler, Claudia Kolmakov, Kirill Wurm, Christian A. Willig, Katrin I. Hell, Stefan W. |
author |
Belov, Vladimir N. |
author_facet |
Belov, Vladimir N. Mitronova, Gyuzel Yu. Bossi, Mariano Luis Boyarskiy, Vadim P. Hebisch, Elke Geisler, Claudia Kolmakov, Kirill Wurm, Christian A. Willig, Katrin I. Hell, Stefan W. |
author_role |
author |
author2 |
Mitronova, Gyuzel Yu. Bossi, Mariano Luis Boyarskiy, Vadim P. Hebisch, Elke Geisler, Claudia Kolmakov, Kirill Wurm, Christian A. Willig, Katrin I. Hell, Stefan W. |
author2_role |
author author author author author author author author author |
dc.subject.none.fl_str_mv |
Bioconjugation Fluorescence Photolysis Rhodamines |
topic |
Bioconjugation Fluorescence Photolysis Rhodamines |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.4 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
Caged rhodamine dyes (Rhodamines NN) of five basic colors were synthesized and used as “hidden” markers in subdiffractional and conventional light microscopy. These masked fluorophores with a 2-diazo-1-indanone group can be irreversibly photoactivated, either by irradiation with UV- or violet light (one-photon process), or by exposure to intense red light (λ∼750 nm; two-photon mode). All dyes possess a very small 2-diazoketone caging group incorporated into the 2-diazo-1-indanone residue with a quaternary carbon atom (C-3) and a spiro-9H-xanthene fragment. Initially they are non-colored (pale yellow), non-fluorescent, and absorb at λ=330–350 nm (molar extinction coefficient (ε)≈104 M−1 cm−1) with a band edge that extends to about λ=440 nm. The absorption and emission bands of the uncaged derivatives are tunable over a wide range (λ=511–633 and 525–653 nm, respectively). The unmasked dyes are highly colored and fluorescent (ε= 3–8×104 M−1 cm−1 and fluorescence quantum yields (ϕ)=40–85 % in the unbound state and in methanol). By stepwise and orthogonal protection of carboxylic and sulfonic acid groups a highly water-soluble caged red-emitting dye with two sulfonic acid residues was prepared. Rhodamines NN were decorated with amino-reactive N-hydroxysuccinimidyl ester groups, applied in aqueous buffers, easily conjugated with proteins, and readily photoactivated (uncaged) with λ=375–420 nm light or intense red light (λ=775 nm). Protein conjugates with optimal degrees of labeling (3–6) were prepared and uncaged with λ=405 nm light in aqueous buffer solutions (ϕ=20–38 %). The photochemical cleavage of the masking group generates only molecular nitrogen. Some 10–40 % of the non-fluorescent (dark) byproducts are also formed. However, they have low absorbance and do not quench the fluorescence of the uncaged dyes. Photoactivation of the individual molecules of Rhodamines NN (e.g., due to reversible or irreversible transition to a “dark” non-emitting state or photobleaching) provides multicolor images with subdiffractional optical resolution. The applicability of these novel caged fluorophores in super-resolution optical microscopy is exemplified. Fil: Belov, Vladimir N.. Max Planck Institute for Biophysical Chemistry ; Alemania Fil: Mitronova, Gyuzel Yu.. Max Planck Institute for Biophysical Chemistry ; Alemania Fil: Bossi, Mariano Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina Fil: Boyarskiy, Vadim P.. St. Petersburg State University; Rusia Fil: Hebisch, Elke. Max Planck Institute for Biophysical Chemistry ; Alemania Fil: Geisler, Claudia. Max Planck Institute for Biophysical Chemistry ; Alemania Fil: Kolmakov, Kirill. Max Planck Institute for Biophysical Chemistry ; Alemania Fil: Wurm, Christian A.. Max Planck Institute for Biophysical Chemistry ; Alemania Fil: Willig, Katrin I.. Max Planck Institute for Biophysical Chemistry ; Alemania Fil: Hell, Stefan W.. Max Planck Institute for Biophysical Chemistry ; Alemania |
description |
Caged rhodamine dyes (Rhodamines NN) of five basic colors were synthesized and used as “hidden” markers in subdiffractional and conventional light microscopy. These masked fluorophores with a 2-diazo-1-indanone group can be irreversibly photoactivated, either by irradiation with UV- or violet light (one-photon process), or by exposure to intense red light (λ∼750 nm; two-photon mode). All dyes possess a very small 2-diazoketone caging group incorporated into the 2-diazo-1-indanone residue with a quaternary carbon atom (C-3) and a spiro-9H-xanthene fragment. Initially they are non-colored (pale yellow), non-fluorescent, and absorb at λ=330–350 nm (molar extinction coefficient (ε)≈104 M−1 cm−1) with a band edge that extends to about λ=440 nm. The absorption and emission bands of the uncaged derivatives are tunable over a wide range (λ=511–633 and 525–653 nm, respectively). The unmasked dyes are highly colored and fluorescent (ε= 3–8×104 M−1 cm−1 and fluorescence quantum yields (ϕ)=40–85 % in the unbound state and in methanol). By stepwise and orthogonal protection of carboxylic and sulfonic acid groups a highly water-soluble caged red-emitting dye with two sulfonic acid residues was prepared. Rhodamines NN were decorated with amino-reactive N-hydroxysuccinimidyl ester groups, applied in aqueous buffers, easily conjugated with proteins, and readily photoactivated (uncaged) with λ=375–420 nm light or intense red light (λ=775 nm). Protein conjugates with optimal degrees of labeling (3–6) were prepared and uncaged with λ=405 nm light in aqueous buffer solutions (ϕ=20–38 %). The photochemical cleavage of the masking group generates only molecular nitrogen. Some 10–40 % of the non-fluorescent (dark) byproducts are also formed. However, they have low absorbance and do not quench the fluorescence of the uncaged dyes. Photoactivation of the individual molecules of Rhodamines NN (e.g., due to reversible or irreversible transition to a “dark” non-emitting state or photobleaching) provides multicolor images with subdiffractional optical resolution. The applicability of these novel caged fluorophores in super-resolution optical microscopy is exemplified. |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-09 |
dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
format |
article |
status_str |
publishedVersion |
dc.identifier.none.fl_str_mv |
http://hdl.handle.net/11336/32179 Belov, Vladimir N.; Hebisch, Elke; Mitronova, Gyuzel Yu.; Bossi, Mariano Luis; Boyarskiy, Vadim P.; Geisler, Claudia; et al.; Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications; Wiley VCH Verlag; Chemistry- A European Journal; 20; 41; 9-2014; 13162-13173 0947-6539 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/32179 |
identifier_str_mv |
Belov, Vladimir N.; Hebisch, Elke; Mitronova, Gyuzel Yu.; Bossi, Mariano Luis; Boyarskiy, Vadim P.; Geisler, Claudia; et al.; Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications; Wiley VCH Verlag; Chemistry- A European Journal; 20; 41; 9-2014; 13162-13173 0947-6539 CONICET Digital CONICET |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/doi/10.1002/chem.201403316 info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1002/chem.201403316/abstract |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Wiley VCH Verlag |
publisher.none.fl_str_mv |
Wiley VCH Verlag |
dc.source.none.fl_str_mv |
reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
reponame_str |
CONICET Digital (CONICET) |
collection |
CONICET Digital (CONICET) |
instname_str |
Consejo Nacional de Investigaciones Científicas y Técnicas |
repository.name.fl_str_mv |
CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
repository.mail.fl_str_mv |
dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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1842269602249179136 |
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13.13397 |