Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes

Autores
Balzarotti, Francisco; Leanza, Yvan; Gwosch, Klaus C.; Gynna, Arvid H.; Westphal, Volker; Stefani, Fernando Daniel; Elf, Johan; Hell, Stefan W.
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
We introduce MINFLUX, a concept for localizing photon emitters in space. By probing the emitter with a local intensity minimum of excitation light, MINFLUX minimizes the fluorescence photons needed for high localization precision. In our experiments, 22 times fewer fluorescence photons are required as compared to popular centroid localization. In superresolution microscopy, MINFLUX attained ~1-nanometer precision, resolving molecules only 6 nanometers apart. MINFLUX tracking of single fluorescent proteins increased the temporal resolution and the number of localizations per trace by a factor of 100, as demonstrated with diffusing 30S ribosomal subunits in living Escherichia coli. As conceptual limits have not been reached, we expect this localization modality to break new ground for observing the dynamics, distribution, and structure of macromolecules in living cells and beyond.
Fil: Balzarotti, Francisco. Max Planck Institute for Biophysical Chemistry; Alemania
Fil: Leanza, Yvan. Max Planck Institute for Biophysical Chemistry; Alemania
Fil: Gwosch, Klaus C.. Max Planck Institute for Biophysical Chemistry; Alemania
Fil: Gynna, Arvid H.. Uppsala University; Suecia
Fil: Westphal, Volker. Max Planck Institute for Biophysical Chemistry; Alemania
Fil: Stefani, Fernando Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias ; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Física; Argentina
Fil: Elf, Johan. Uppsala University; Suecia
Fil: Hell, Stefan W.. Max Planck Institute for Biophysical Chemistry; Alemania. German Cancer Research Center; Alemania
Materia
SUPERRESOLUTION
FLUORESCENCE MICROSCOPY
BIOIMAGING
PHOTON BUDGET
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:11336/54290
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxesBalzarotti, FranciscoLeanza, YvanGwosch, Klaus C.Gynna, Arvid H.Westphal, VolkerStefani, Fernando DanielElf, JohanHell, Stefan W.SUPERRESOLUTIONFLUORESCENCE MICROSCOPYBIOIMAGINGPHOTON BUDGEThttps://purl.org/becyt/ford/1.3https://purl.org/becyt/ford/1We introduce MINFLUX, a concept for localizing photon emitters in space. By probing the emitter with a local intensity minimum of excitation light, MINFLUX minimizes the fluorescence photons needed for high localization precision. In our experiments, 22 times fewer fluorescence photons are required as compared to popular centroid localization. In superresolution microscopy, MINFLUX attained ~1-nanometer precision, resolving molecules only 6 nanometers apart. MINFLUX tracking of single fluorescent proteins increased the temporal resolution and the number of localizations per trace by a factor of 100, as demonstrated with diffusing 30S ribosomal subunits in living Escherichia coli. As conceptual limits have not been reached, we expect this localization modality to break new ground for observing the dynamics, distribution, and structure of macromolecules in living cells and beyond.Fil: Balzarotti, Francisco. Max Planck Institute for Biophysical Chemistry; AlemaniaFil: Leanza, Yvan. Max Planck Institute for Biophysical Chemistry; AlemaniaFil: Gwosch, Klaus C.. Max Planck Institute for Biophysical Chemistry; AlemaniaFil: Gynna, Arvid H.. Uppsala University; SueciaFil: Westphal, Volker. Max Planck Institute for Biophysical Chemistry; AlemaniaFil: Stefani, Fernando Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias ; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Física; ArgentinaFil: Elf, Johan. Uppsala University; SueciaFil: Hell, Stefan W.. Max Planck Institute for Biophysical Chemistry; Alemania. German Cancer Research Center; AlemaniaAmerican Association for the Advancement of Science2017-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/54290Balzarotti, Francisco; Leanza, Yvan; Gwosch, Klaus C.; Gynna, Arvid H.; Westphal, Volker; et al.; Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes; American Association for the Advancement of Science; Science; 355; 6325; 2-2017; 606-6120036-80751095-9203CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1126/science.aak9913info:eu-repo/semantics/altIdentifier/url/https://arxiv.org/abs/1611.03401info:eu-repo/semantics/altIdentifier/url/http://science.sciencemag.org/content/355/6325/606info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:43:33Zoai:ri.conicet.gov.ar:11336/54290instacron: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 15:43:34.014CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes
title Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes
spellingShingle Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes
Balzarotti, Francisco
SUPERRESOLUTION
FLUORESCENCE MICROSCOPY
BIOIMAGING
PHOTON BUDGET
title_short Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes
title_full Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes
title_fullStr Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes
title_full_unstemmed Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes
title_sort Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes
dc.creator.none.fl_str_mv Balzarotti, Francisco
Leanza, Yvan
Gwosch, Klaus C.
Gynna, Arvid H.
Westphal, Volker
Stefani, Fernando Daniel
Elf, Johan
Hell, Stefan W.
author Balzarotti, Francisco
author_facet Balzarotti, Francisco
Leanza, Yvan
Gwosch, Klaus C.
Gynna, Arvid H.
Westphal, Volker
Stefani, Fernando Daniel
Elf, Johan
Hell, Stefan W.
author_role author
author2 Leanza, Yvan
Gwosch, Klaus C.
Gynna, Arvid H.
Westphal, Volker
Stefani, Fernando Daniel
Elf, Johan
Hell, Stefan W.
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv SUPERRESOLUTION
FLUORESCENCE MICROSCOPY
BIOIMAGING
PHOTON BUDGET
topic SUPERRESOLUTION
FLUORESCENCE MICROSCOPY
BIOIMAGING
PHOTON BUDGET
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.3
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv We introduce MINFLUX, a concept for localizing photon emitters in space. By probing the emitter with a local intensity minimum of excitation light, MINFLUX minimizes the fluorescence photons needed for high localization precision. In our experiments, 22 times fewer fluorescence photons are required as compared to popular centroid localization. In superresolution microscopy, MINFLUX attained ~1-nanometer precision, resolving molecules only 6 nanometers apart. MINFLUX tracking of single fluorescent proteins increased the temporal resolution and the number of localizations per trace by a factor of 100, as demonstrated with diffusing 30S ribosomal subunits in living Escherichia coli. As conceptual limits have not been reached, we expect this localization modality to break new ground for observing the dynamics, distribution, and structure of macromolecules in living cells and beyond.
Fil: Balzarotti, Francisco. Max Planck Institute for Biophysical Chemistry; Alemania
Fil: Leanza, Yvan. Max Planck Institute for Biophysical Chemistry; Alemania
Fil: Gwosch, Klaus C.. Max Planck Institute for Biophysical Chemistry; Alemania
Fil: Gynna, Arvid H.. Uppsala University; Suecia
Fil: Westphal, Volker. Max Planck Institute for Biophysical Chemistry; Alemania
Fil: Stefani, Fernando Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias ; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Física; Argentina
Fil: Elf, Johan. Uppsala University; Suecia
Fil: Hell, Stefan W.. Max Planck Institute for Biophysical Chemistry; Alemania. German Cancer Research Center; Alemania
description We introduce MINFLUX, a concept for localizing photon emitters in space. By probing the emitter with a local intensity minimum of excitation light, MINFLUX minimizes the fluorescence photons needed for high localization precision. In our experiments, 22 times fewer fluorescence photons are required as compared to popular centroid localization. In superresolution microscopy, MINFLUX attained ~1-nanometer precision, resolving molecules only 6 nanometers apart. MINFLUX tracking of single fluorescent proteins increased the temporal resolution and the number of localizations per trace by a factor of 100, as demonstrated with diffusing 30S ribosomal subunits in living Escherichia coli. As conceptual limits have not been reached, we expect this localization modality to break new ground for observing the dynamics, distribution, and structure of macromolecules in living cells and beyond.
publishDate 2017
dc.date.none.fl_str_mv 2017-02
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/11336/54290
Balzarotti, Francisco; Leanza, Yvan; Gwosch, Klaus C.; Gynna, Arvid H.; Westphal, Volker; et al.; Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes; American Association for the Advancement of Science; Science; 355; 6325; 2-2017; 606-612
0036-8075
1095-9203
CONICET Digital
CONICET
url http://hdl.handle.net/11336/54290
identifier_str_mv Balzarotti, Francisco; Leanza, Yvan; Gwosch, Klaus C.; Gynna, Arvid H.; Westphal, Volker; et al.; Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes; American Association for the Advancement of Science; Science; 355; 6325; 2-2017; 606-612
0036-8075
1095-9203
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1126/science.aak9913
info:eu-repo/semantics/altIdentifier/url/https://arxiv.org/abs/1611.03401
info:eu-repo/semantics/altIdentifier/url/http://science.sciencemag.org/content/355/6325/606
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
application/pdf
dc.publisher.none.fl_str_mv American Association for the Advancement of Science
publisher.none.fl_str_mv American Association for the Advancement of Science
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_ 1846083540466270208
score 13.22299

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