Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy

Autores
Gensch, Thomas; Bohmer, Martin Federico; Aramendia, Pedro Francisco
Año de publicación
2005
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Single molecule fluorescence detection of Atto590 in poly(vinyl alcohol) was achieved by using a wide-field epifluorescence microscope with CCD-camera detection. Image sequences are obtained from which the time traces of the detected molecules are built. We find a distinctive difference between the time evolution of the fluorescence originating from the molecules detected in the first image of the sequence compared to the time evolution of the fluorescence of the molecules detected in each image of the sequence. Atto590 shows very long blinking times and photobleaching and photoblinking that are both quadratically dependent on the irradiation power density. Our approach allows kinetic separation of photobleaching from blinking. The possibility of choosing different ensembles of molecules is demonstrated and taken advantage of for this aim. Initially dark molecules or low emitting ones that might be overlooked are important to describe the complete ensemble behavior.
Fil: Gensch, Thomas. Research Centre Julich GmbH; Alemania
Fil: Bohmer, Martin Federico. Research Centre Julich GmbH; Alemania
Fil: Aramendia, Pedro Francisco. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Inorgánica, Analítica y Química Física; Argentina. 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
Materia
single molecule
blinking
atto590
fluorescence
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/103415
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/103415
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence MicroscopyGensch, ThomasBohmer, Martin FedericoAramendia, Pedro Franciscosingle moleculeblinkingatto590fluorescencehttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Single molecule fluorescence detection of Atto590 in poly(vinyl alcohol) was achieved by using a wide-field epifluorescence microscope with CCD-camera detection. Image sequences are obtained from which the time traces of the detected molecules are built. We find a distinctive difference between the time evolution of the fluorescence originating from the molecules detected in the first image of the sequence compared to the time evolution of the fluorescence of the molecules detected in each image of the sequence. Atto590 shows very long blinking times and photobleaching and photoblinking that are both quadratically dependent on the irradiation power density. Our approach allows kinetic separation of photobleaching from blinking. The possibility of choosing different ensembles of molecules is demonstrated and taken advantage of for this aim. Initially dark molecules or low emitting ones that might be overlooked are important to describe the complete ensemble behavior.Fil: Gensch, Thomas. Research Centre Julich GmbH; AlemaniaFil: Bohmer, Martin Federico. Research Centre Julich GmbH; AlemaniaFil: Aramendia, Pedro Francisco. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Inorgánica, Analítica y Química Física; Argentina. 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; ArgentinaAmerican Chemical Society2005-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/103415Gensch, Thomas; Bohmer, Martin Federico; Aramendia, Pedro Francisco; Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy; American Chemical Society; Journal of Physical Chemistry A; 109; 30; 8-2005; 6652-66581089-5639CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1021/jp0510847info:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/10.1021/jp0510847info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:22:53Zoai:ri.conicet.gov.ar:11336/103415instacron: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:34982025-10-15 15:22:54.228CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy
title Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy
spellingShingle Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy
Gensch, Thomas
single molecule
blinking
atto590
fluorescence
title_short Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy
title_full Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy
title_fullStr Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy
title_full_unstemmed Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy
title_sort Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy
dc.creator.none.fl_str_mv Gensch, Thomas
Bohmer, Martin Federico
Aramendia, Pedro Francisco
author Gensch, Thomas
author_facet Gensch, Thomas
Bohmer, Martin Federico
Aramendia, Pedro Francisco
author_role author
author2 Bohmer, Martin Federico
Aramendia, Pedro Francisco
author2_role author
author
dc.subject.none.fl_str_mv single molecule
blinking
atto590
fluorescence
topic single molecule
blinking
atto590
fluorescence
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Single molecule fluorescence detection of Atto590 in poly(vinyl alcohol) was achieved by using a wide-field epifluorescence microscope with CCD-camera detection. Image sequences are obtained from which the time traces of the detected molecules are built. We find a distinctive difference between the time evolution of the fluorescence originating from the molecules detected in the first image of the sequence compared to the time evolution of the fluorescence of the molecules detected in each image of the sequence. Atto590 shows very long blinking times and photobleaching and photoblinking that are both quadratically dependent on the irradiation power density. Our approach allows kinetic separation of photobleaching from blinking. The possibility of choosing different ensembles of molecules is demonstrated and taken advantage of for this aim. Initially dark molecules or low emitting ones that might be overlooked are important to describe the complete ensemble behavior.
Fil: Gensch, Thomas. Research Centre Julich GmbH; Alemania
Fil: Bohmer, Martin Federico. Research Centre Julich GmbH; Alemania
Fil: Aramendia, Pedro Francisco. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Inorgánica, Analítica y Química Física; Argentina. 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
description Single molecule fluorescence detection of Atto590 in poly(vinyl alcohol) was achieved by using a wide-field epifluorescence microscope with CCD-camera detection. Image sequences are obtained from which the time traces of the detected molecules are built. We find a distinctive difference between the time evolution of the fluorescence originating from the molecules detected in the first image of the sequence compared to the time evolution of the fluorescence of the molecules detected in each image of the sequence. Atto590 shows very long blinking times and photobleaching and photoblinking that are both quadratically dependent on the irradiation power density. Our approach allows kinetic separation of photobleaching from blinking. The possibility of choosing different ensembles of molecules is demonstrated and taken advantage of for this aim. Initially dark molecules or low emitting ones that might be overlooked are important to describe the complete ensemble behavior.
publishDate 2005
dc.date.none.fl_str_mv 2005-08
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/11336/103415
Gensch, Thomas; Bohmer, Martin Federico; Aramendia, Pedro Francisco; Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy; American Chemical Society; Journal of Physical Chemistry A; 109; 30; 8-2005; 6652-6658
1089-5639
CONICET Digital
CONICET
url http://hdl.handle.net/11336/103415
identifier_str_mv Gensch, Thomas; Bohmer, Martin Federico; Aramendia, Pedro Francisco; Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy; American Chemical Society; Journal of Physical Chemistry A; 109; 30; 8-2005; 6652-6658
1089-5639
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1021/jp0510847
info:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/10.1021/jp0510847
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 American Chemical Society
publisher.none.fl_str_mv American Chemical Society
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
_version_ 1846083376163848192
score 13.22299

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