Magnetic alignment of rhodamine/magnetite dual-labeled microtubules probed with inverted fluorescence microscopy
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Anais da Academia Brasileira de Ciências (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652022000500502 |
Resumo: | Abstract Molecular machines, as exemplified by the kinesin and microtubule system, are responsible for molecular transport in cells. The monitoring of the cellular machinery has attracted much attention in recent years, requiring sophisticated techniques such as optical tweezers, and dark field hyperspectral and fluorescence microscopies. It also demands suitable procedures for immobilization and labeling with functional agents such as dyes, plasmonic nanoparticles and quantum dots. In this work, microtubules were co-polymerized by incubating a tubulin mix consisting of 7 biotinylated tubulin to 3 rhodamine tubulin. Rhodamine provided the fluorescent tag, while biotin was the anchoring group for receiving streptavidin containing species. To control the microtubule alignment and consequently, the molecular gliding directions, functionalized iron oxide nanoparticles were employed in the presence of an external magnet field. Such iron oxide nanoparticles, (MagNPs) were previously coated with silica and (3-aminopro-pyl)triethoxysilane (APTS) and then modified with streptavidin (SA) for linking to the biotin-functionalized microtubules. In this way, the binding has been successfully performed, and the magnetic alignment probed by Inverted Fluorescence Microscopy. The proposed strategy has proved promising, as tested with one of the most important biological structures of the cellular machinery. |
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Magnetic alignment of rhodamine/magnetite dual-labeled microtubules probed with inverted fluorescence microscopyMolecular machineskinesin and microtubulesmagnetic nanoparticlesfluorescence microscopyCytoViva hypermicroscopyAbstract Molecular machines, as exemplified by the kinesin and microtubule system, are responsible for molecular transport in cells. The monitoring of the cellular machinery has attracted much attention in recent years, requiring sophisticated techniques such as optical tweezers, and dark field hyperspectral and fluorescence microscopies. It also demands suitable procedures for immobilization and labeling with functional agents such as dyes, plasmonic nanoparticles and quantum dots. In this work, microtubules were co-polymerized by incubating a tubulin mix consisting of 7 biotinylated tubulin to 3 rhodamine tubulin. Rhodamine provided the fluorescent tag, while biotin was the anchoring group for receiving streptavidin containing species. To control the microtubule alignment and consequently, the molecular gliding directions, functionalized iron oxide nanoparticles were employed in the presence of an external magnet field. Such iron oxide nanoparticles, (MagNPs) were previously coated with silica and (3-aminopro-pyl)triethoxysilane (APTS) and then modified with streptavidin (SA) for linking to the biotin-functionalized microtubules. In this way, the binding has been successfully performed, and the magnetic alignment probed by Inverted Fluorescence Microscopy. The proposed strategy has proved promising, as tested with one of the most important biological structures of the cellular machinery.Academia Brasileira de Ciências2022-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652022000500502Anais da Academia Brasileira de Ciências v.94 n.3 2022reponame:Anais da Academia Brasileira de Ciências (Online)instname:Academia Brasileira de Ciências (ABC)instacron:ABC10.1590/0001-3765202220210917info:eu-repo/semantics/openAccessTOMA,HENRIQUE EISIOLIVEIRA,DANIELMELO,FERNANDO M. DEeng2022-07-28T00:00:00Zoai:scielo:S0001-37652022000500502Revistahttp://www.scielo.br/aabchttps://old.scielo.br/oai/scielo-oai.php||aabc@abc.org.br1678-26900001-3765opendoar:2022-07-28T00:00Anais da Academia Brasileira de Ciências (Online) - Academia Brasileira de Ciências (ABC)false |
dc.title.none.fl_str_mv |
Magnetic alignment of rhodamine/magnetite dual-labeled microtubules probed with inverted fluorescence microscopy |
title |
Magnetic alignment of rhodamine/magnetite dual-labeled microtubules probed with inverted fluorescence microscopy |
spellingShingle |
Magnetic alignment of rhodamine/magnetite dual-labeled microtubules probed with inverted fluorescence microscopy TOMA,HENRIQUE EISI Molecular machines kinesin and microtubules magnetic nanoparticles fluorescence microscopy CytoViva hypermicroscopy |
title_short |
Magnetic alignment of rhodamine/magnetite dual-labeled microtubules probed with inverted fluorescence microscopy |
title_full |
Magnetic alignment of rhodamine/magnetite dual-labeled microtubules probed with inverted fluorescence microscopy |
title_fullStr |
Magnetic alignment of rhodamine/magnetite dual-labeled microtubules probed with inverted fluorescence microscopy |
title_full_unstemmed |
Magnetic alignment of rhodamine/magnetite dual-labeled microtubules probed with inverted fluorescence microscopy |
title_sort |
Magnetic alignment of rhodamine/magnetite dual-labeled microtubules probed with inverted fluorescence microscopy |
author |
TOMA,HENRIQUE EISI |
author_facet |
TOMA,HENRIQUE EISI OLIVEIRA,DANIEL MELO,FERNANDO M. DE |
author_role |
author |
author2 |
OLIVEIRA,DANIEL MELO,FERNANDO M. DE |
author2_role |
author author |
dc.contributor.author.fl_str_mv |
TOMA,HENRIQUE EISI OLIVEIRA,DANIEL MELO,FERNANDO M. DE |
dc.subject.por.fl_str_mv |
Molecular machines kinesin and microtubules magnetic nanoparticles fluorescence microscopy CytoViva hypermicroscopy |
topic |
Molecular machines kinesin and microtubules magnetic nanoparticles fluorescence microscopy CytoViva hypermicroscopy |
description |
Abstract Molecular machines, as exemplified by the kinesin and microtubule system, are responsible for molecular transport in cells. The monitoring of the cellular machinery has attracted much attention in recent years, requiring sophisticated techniques such as optical tweezers, and dark field hyperspectral and fluorescence microscopies. It also demands suitable procedures for immobilization and labeling with functional agents such as dyes, plasmonic nanoparticles and quantum dots. In this work, microtubules were co-polymerized by incubating a tubulin mix consisting of 7 biotinylated tubulin to 3 rhodamine tubulin. Rhodamine provided the fluorescent tag, while biotin was the anchoring group for receiving streptavidin containing species. To control the microtubule alignment and consequently, the molecular gliding directions, functionalized iron oxide nanoparticles were employed in the presence of an external magnet field. Such iron oxide nanoparticles, (MagNPs) were previously coated with silica and (3-aminopro-pyl)triethoxysilane (APTS) and then modified with streptavidin (SA) for linking to the biotin-functionalized microtubules. In this way, the binding has been successfully performed, and the magnetic alignment probed by Inverted Fluorescence Microscopy. The proposed strategy has proved promising, as tested with one of the most important biological structures of the cellular machinery. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-01-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652022000500502 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652022000500502 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/0001-3765202220210917 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Academia Brasileira de Ciências |
publisher.none.fl_str_mv |
Academia Brasileira de Ciências |
dc.source.none.fl_str_mv |
Anais da Academia Brasileira de Ciências v.94 n.3 2022 reponame:Anais da Academia Brasileira de Ciências (Online) instname:Academia Brasileira de Ciências (ABC) instacron:ABC |
instname_str |
Academia Brasileira de Ciências (ABC) |
instacron_str |
ABC |
institution |
ABC |
reponame_str |
Anais da Academia Brasileira de Ciências (Online) |
collection |
Anais da Academia Brasileira de Ciências (Online) |
repository.name.fl_str_mv |
Anais da Academia Brasileira de Ciências (Online) - Academia Brasileira de Ciências (ABC) |
repository.mail.fl_str_mv |
||aabc@abc.org.br |
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1754302872103682048 |