TY - JOUR T1 - Microtubule organization within mitotic spindles revealed by serial block face scanning EM and image analysis JF - bioRxiv DO - 10.1101/087866 SP - 087866 AU - Faye M. Nixon AU - Thomas R. Honnor AU - Georgina P. Starling AU - Alison J. Beckett AU - Adam M. Johansen AU - Julia A. Brettschneider AU - Ian A. Prior AU - Stephen J. Royle Y1 - 2016/01/01 UR - http://biorxiv.org/content/early/2016/11/15/087866.abstract N2 - 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. ER -