Important factors determining the nanoscale tracking precision of dynamic microtubule ends
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- dc.contributor.author Bohner, G.ca
- dc.contributor.author Gustafsson, N.ca
- dc.contributor.author Cade, Nicholas Ianca
- dc.contributor.author Maurer, Sebastianca
- dc.contributor.author Griffin, L. D.ca
- dc.contributor.author Surrey, Thomasca
- dc.date.accessioned 2017-04-24T10:29:33Z
- dc.date.available 2017-04-24T10:29:33Z
- dc.date.issued 2016
- dc.description.abstract 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.
- dc.format.mimetype application/pdfca
- dc.identifier.citation Bohner G, Gustafsson N, Cade NI, Maurer S, Griffin LD, Surrey T. Important factors determining the nanoscale tracking precision of dynamic microtubule ends. Journal of Microscopy. 2016; 261(1): 67-78. DOI: 10.1111/jmi.12316
- dc.identifier.doi http://dx.doi.org/10.1111/jmi.12316
- dc.identifier.issn 0022-2720
- dc.identifier.uri http://hdl.handle.net/10230/30880
- dc.language.iso eng
- dc.publisher Wileyca
- dc.relation.ispartof Journal of Microscopy. 2016; 261(1): 67-78
- dc.rights © 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.
- dc.rights.accessRights info:eu-repo/semantics/openAccess
- dc.rights.uri https://creativecommons.org/licenses/by/4.0/
- dc.subject.keyword Image processing
- dc.subject.keyword Image simulation
- dc.subject.keyword Microtubule
- dc.subject.keyword TIRF microscopy
- dc.subject.keyword Tracking
- dc.title Important factors determining the nanoscale tracking precision of dynamic microtubule endsca
- dc.type info:eu-repo/semantics/article
- dc.type.version info:eu-repo/semantics/publishedVersion