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Microtubule organization within mitotic spindles revealed by serial block face scanning electron microscopy and image analysis
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Nixon, Faye M., Honnor, Thomas R., Clarke, Nicholas I., Starling, Georgina, Beckett, Alison J., Johansen, Adam M., Brettschneider, Julia, Prior, Ian A. and Royle, Stephen J. (2017) Microtubule organization within mitotic spindles revealed by serial block face scanning electron microscopy and image analysis. Journal of Cell Science, 130 (10). pp. 1845-1855. doi:10.1242/jcs.203877 ISSN 0021-9533.
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Official URL: http://dx.doi.org/10.1242/jcs.203877
Abstract
Serial block face scanning electron microscopy (SBF-SEM) is a powerful method to analyze cells in 3D. Here, working at the resolution limit of the method, we describe a correlative light–SBF-SEM workflow to resolve microtubules of the mitotic spindle in human cells. We present four examples of uses for this workflow that are not practical by light microscopy and/or transmission electron microscopy. First, distinguishing closely associated microtubules within K-fibers; second, resolving bridging fibers in the mitotic spindle; third, visualizing membranes in mitotic cells, relative to the spindle apparatus; and fourth, volumetric analysis of kinetochores. Our workflow also includes new computational tools for exploring the spatial arrangement of microtubules within the mitotic spindle. We use these tools to show that microtubule order in mitotic spindles is sensitive to the level of TACC3 on the spindle.
Item Type: | Journal Article | ||||||||
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Subjects: | Q Science > QH Natural history > QH301 Biology | ||||||||
Divisions: | Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences > Cell & Developmental Biology Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences Faculty of Science, Engineering and Medicine > Science > Statistics Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School |
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Library of Congress Subject Headings (LCSH): | Electron microscopy -- Technique, Microtubules, Mitosis, Cell cycle | ||||||||
Journal or Publication Title: | Journal of Cell Science | ||||||||
Publisher: | The Company of Biologists Ltd. | ||||||||
ISSN: | 0021-9533 | ||||||||
Official Date: | 15 May 2017 | ||||||||
Dates: |
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Volume: | 130 | ||||||||
Number: | 10 | ||||||||
Page Range: | pp. 1845-1855 | ||||||||
DOI: | 10.1242/jcs.203877 | ||||||||
Status: | Peer Reviewed | ||||||||
Publication Status: | Published | ||||||||
Access rights to Published version: | Restricted or Subscription Access | ||||||||
Date of first compliant deposit: | 2 October 2017 | ||||||||
Date of first compliant Open Access: | 15 May 2018 | ||||||||
Funder: | North West Cancer Research, Cancer Research UK (CRUK) | ||||||||
Grant number: | C25425/A15182 (CRUK) | ||||||||
Related URLs: | |||||||||
Open Access Version: |
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