Important factors determining the nanoscale tracking precision of dynamic microtubule ends

Tracking dynamic microtubule ends in fluorescence microscopy movies provides insight into the statistical properties of microtubule dynamics and is vital for further analysis that requires knowledge of the trajectories of the microtubule ends. Here we analyse the performance of a previously develope...

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Detalhes bibliográficos
Autores: Bohner, G., Gustafsson, N., Cade, Nicholas Ian, Maurer, Sebastian, Griffin, L. D., Surrey, Thomas
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2016
País:España
Recursos:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10230/30880
Acesso em linha:http://hdl.handle.net/10230/30880
http://dx.doi.org/10.1111/jmi.12316
Access Level:acceso abierto
Palavra-chave:Image processing
Image simulation
Microtubule
TIRF microscopy
Tracking
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spelling Important factors determining the nanoscale tracking precision of dynamic microtubule endsBohner, G.Gustafsson, N.Cade, Nicholas IanMaurer, SebastianGriffin, L. D.Surrey, ThomasImage processingImage simulationMicrotubuleTIRF microscopyTrackingTracking dynamic microtubule ends in fluorescence microscopy movies provides insight into the statistical properties of microtubule dynamics and is vital for further analysis that requires knowledge of the trajectories of the microtubule ends. Here we analyse the performance of a previously developed automated microtubule end tracking routine; this has been optimized for comparatively low signal-to-noise image sequences that are characteristic of microscopy movies of dynamic microtubules growing in vitro. Sequences of simulated microtubule images were generated assuming a variety of different experimental conditions. The simulated movies were then tracked and the tracking errors were characterized. We found that the growth characteristics of the microtubules within realistic ranges had a negligible effect on the tracking precision. The fluorophore labelling density, the pixel size of the images, and the exposure times were found to be important parameters limiting the tracking precision which could be explained using concepts of single molecule localization microscopy. The signal-to-noise ratio was found to be a good single predictor of the tracking precision: typical experimental signal-to-noise ratios lead to tracking precisions in the range of tens of nanometres, making the tracking program described here a useful tool for dynamic microtubule end tracking with close to molecular precision.Wiley201720172016info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/10230/30880http://dx.doi.org/10.1111/jmi.12316reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésJournal of Microscopy. 2016; 261(1): 67-78© 2015 The Authors. Journal of Microscopy published by John Wiley & Sons. Ltd on behalf of Royal Microscopical Society. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.https://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:recercat.cat:10230/308802026-05-29T05:05:01Z
dc.title.none.fl_str_mv Important factors determining the nanoscale tracking precision of dynamic microtubule ends
title Important factors determining the nanoscale tracking precision of dynamic microtubule ends
spellingShingle Important factors determining the nanoscale tracking precision of dynamic microtubule ends
Bohner, G.
Image processing
Image simulation
Microtubule
TIRF microscopy
Tracking
title_short Important factors determining the nanoscale tracking precision of dynamic microtubule ends
title_full Important factors determining the nanoscale tracking precision of dynamic microtubule ends
title_fullStr Important factors determining the nanoscale tracking precision of dynamic microtubule ends
title_full_unstemmed Important factors determining the nanoscale tracking precision of dynamic microtubule ends
title_sort Important factors determining the nanoscale tracking precision of dynamic microtubule ends
dc.creator.none.fl_str_mv Bohner, G.
Gustafsson, N.
Cade, Nicholas Ian
Maurer, Sebastian
Griffin, L. D.
Surrey, Thomas
author Bohner, G.
author_facet Bohner, G.
Gustafsson, N.
Cade, Nicholas Ian
Maurer, Sebastian
Griffin, L. D.
Surrey, Thomas
author_role author
author2 Gustafsson, N.
Cade, Nicholas Ian
Maurer, Sebastian
Griffin, L. D.
Surrey, Thomas
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Image processing
Image simulation
Microtubule
TIRF microscopy
Tracking
topic Image processing
Image simulation
Microtubule
TIRF microscopy
Tracking
description Tracking dynamic microtubule ends in fluorescence microscopy movies provides insight into the statistical properties of microtubule dynamics and is vital for further analysis that requires knowledge of the trajectories of the microtubule ends. Here we analyse the performance of a previously developed automated microtubule end tracking routine; this has been optimized for comparatively low signal-to-noise image sequences that are characteristic of microscopy movies of dynamic microtubules growing in vitro. Sequences of simulated microtubule images were generated assuming a variety of different experimental conditions. The simulated movies were then tracked and the tracking errors were characterized. We found that the growth characteristics of the microtubules within realistic ranges had a negligible effect on the tracking precision. The fluorophore labelling density, the pixel size of the images, and the exposure times were found to be important parameters limiting the tracking precision which could be explained using concepts of single molecule localization microscopy. The signal-to-noise ratio was found to be a good single predictor of the tracking precision: typical experimental signal-to-noise ratios lead to tracking precisions in the range of tens of nanometres, making the tracking program described here a useful tool for dynamic microtubule end tracking with close to molecular precision.
publishDate 2016
dc.date.none.fl_str_mv 2016
2017
2017
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10230/30880
http://dx.doi.org/10.1111/jmi.12316
url http://hdl.handle.net/10230/30880
http://dx.doi.org/10.1111/jmi.12316
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Journal of Microscopy. 2016; 261(1): 67-78
dc.rights.none.fl_str_mv https://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
dc.source.none.fl_str_mv reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,81155

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