RT Journal Article
SR Electronic
T1 Microtubule organization within mitotic spindles revealed by serial block face scanning EM and image analysis
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 087866
DO 10.1101/087866
A1 Faye M. Nixon
A1 Thomas R. Honnor
A1 Georgina P. Starling
A1 Alison J. Beckett
A1 Adam M. Johansen
A1 Julia A. Brettschneider
A1 Ian A. Prior
A1 Stephen J. Royle
YR 2016
UL http://biorxiv.org/content/early/2016/11/15/087866.abstract
AB 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 three examples of uses for this workflow which are not practical by light microscopy and/or TEM. 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. Our workflow also includes new computational tools for exploring the spatial arrangement of MTs 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.