Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins

Autores
Barrantes, Francisco Jose
Año de publicación
2016
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Recent years have witnessed huge progress in the field of light microscopy with the development and implementation of new approaches leading to dramatic improvements in the spatial and temporal resolution of this form of imaging, most particularly in its biological applications. The limitations in spatial resolution imposed by the diffraction of light have been circumvented by resorting to different strategies, which are briefly outlined in the Introduction. These protocols are intended to provide practical guidelines for the imaging of synaptic proteins using one such strategy, namely, single-molecule stochastic localization super-resolution microscopy. The protocols use neuronal cells from the hippocampus of rodent embryos as the experimental paradigm and outline the steps for obtaining dissociated neurons and establishing primary cultures for in vitro studies. The techniques can be adapted to the culture of neurons from other brain regions. Procedures for handling fixed and live specimens are described, as well as the use of extrinsic fluorescent probes and fluorescent proteins, mounting media, examples of hardware configurations, software for image analysis, and some hints for the implementation of minimalist approaches to single-molecule localization nanoscopy.
Fil: Barrantes, Francisco Jose. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentina
Materia
Nanoscopy
Neuronal Cell Culture
Single-Molecule Imaging
Super-Resolution Microscopy
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:11336/39361
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/39361
network_acronym_str CONICETDig
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network_name_str CONICET Digital (CONICET)
spelling Single-Molecule Localization Super-Resolution Microscopy of Synaptic ProteinsBarrantes, Francisco JoseNanoscopyNeuronal Cell CultureSingle-Molecule ImagingSuper-Resolution Microscopyhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Recent years have witnessed huge progress in the field of light microscopy with the development and implementation of new approaches leading to dramatic improvements in the spatial and temporal resolution of this form of imaging, most particularly in its biological applications. The limitations in spatial resolution imposed by the diffraction of light have been circumvented by resorting to different strategies, which are briefly outlined in the Introduction. These protocols are intended to provide practical guidelines for the imaging of synaptic proteins using one such strategy, namely, single-molecule stochastic localization super-resolution microscopy. The protocols use neuronal cells from the hippocampus of rodent embryos as the experimental paradigm and outline the steps for obtaining dissociated neurons and establishing primary cultures for in vitro studies. The techniques can be adapted to the culture of neurons from other brain regions. Procedures for handling fixed and live specimens are described, as well as the use of extrinsic fluorescent probes and fluorescent proteins, mounting media, examples of hardware configurations, software for image analysis, and some hints for the implementation of minimalist approaches to single-molecule localization nanoscopy.Fil: Barrantes, Francisco Jose. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaHumana Press2016-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/39361Barrantes, Francisco Jose; Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins; Humana Press; Springer Protocols Handbooks; 10-2016; 157-198978-1-4939-6836-71949-2448CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1007/8623_2016_10info:eu-repo/semantics/altIdentifier/url/https://link.springer.com/protocol/10.1007%2F8623_2016_10info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:27:57Zoai:ri.conicet.gov.ar:11336/39361instacron: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 14:27:57.529CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins
title Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins
spellingShingle Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins
Barrantes, Francisco Jose
Nanoscopy
Neuronal Cell Culture
Single-Molecule Imaging
Super-Resolution Microscopy
title_short Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins
title_full Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins
title_fullStr Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins
title_full_unstemmed Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins
title_sort Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins
dc.creator.none.fl_str_mv Barrantes, Francisco Jose
author Barrantes, Francisco Jose
author_facet Barrantes, Francisco Jose
author_role author
dc.subject.none.fl_str_mv Nanoscopy
Neuronal Cell Culture
Single-Molecule Imaging
Super-Resolution Microscopy
topic Nanoscopy
Neuronal Cell Culture
Single-Molecule Imaging
Super-Resolution Microscopy
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Recent years have witnessed huge progress in the field of light microscopy with the development and implementation of new approaches leading to dramatic improvements in the spatial and temporal resolution of this form of imaging, most particularly in its biological applications. The limitations in spatial resolution imposed by the diffraction of light have been circumvented by resorting to different strategies, which are briefly outlined in the Introduction. These protocols are intended to provide practical guidelines for the imaging of synaptic proteins using one such strategy, namely, single-molecule stochastic localization super-resolution microscopy. The protocols use neuronal cells from the hippocampus of rodent embryos as the experimental paradigm and outline the steps for obtaining dissociated neurons and establishing primary cultures for in vitro studies. The techniques can be adapted to the culture of neurons from other brain regions. Procedures for handling fixed and live specimens are described, as well as the use of extrinsic fluorescent probes and fluorescent proteins, mounting media, examples of hardware configurations, software for image analysis, and some hints for the implementation of minimalist approaches to single-molecule localization nanoscopy.
Fil: Barrantes, Francisco Jose. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentina
description Recent years have witnessed huge progress in the field of light microscopy with the development and implementation of new approaches leading to dramatic improvements in the spatial and temporal resolution of this form of imaging, most particularly in its biological applications. The limitations in spatial resolution imposed by the diffraction of light have been circumvented by resorting to different strategies, which are briefly outlined in the Introduction. These protocols are intended to provide practical guidelines for the imaging of synaptic proteins using one such strategy, namely, single-molecule stochastic localization super-resolution microscopy. The protocols use neuronal cells from the hippocampus of rodent embryos as the experimental paradigm and outline the steps for obtaining dissociated neurons and establishing primary cultures for in vitro studies. The techniques can be adapted to the culture of neurons from other brain regions. Procedures for handling fixed and live specimens are described, as well as the use of extrinsic fluorescent probes and fluorescent proteins, mounting media, examples of hardware configurations, software for image analysis, and some hints for the implementation of minimalist approaches to single-molecule localization nanoscopy.
publishDate 2016
dc.date.none.fl_str_mv 2016-10
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/11336/39361
Barrantes, Francisco Jose; Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins; Humana Press; Springer Protocols Handbooks; 10-2016; 157-198
978-1-4939-6836-7
1949-2448
CONICET Digital
CONICET
url http://hdl.handle.net/11336/39361
identifier_str_mv Barrantes, Francisco Jose; Single-Molecule Localization Super-Resolution Microscopy of Synaptic Proteins; Humana Press; Springer Protocols Handbooks; 10-2016; 157-198
978-1-4939-6836-7
1949-2448
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1007/8623_2016_10
info:eu-repo/semantics/altIdentifier/url/https://link.springer.com/protocol/10.1007%2F8623_2016_10
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 Humana Press
publisher.none.fl_str_mv Humana Press
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_ 1846082739598524416
score 13.22299

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