Biology Reference
In-Depth Information
CHAPTER
20
Studying Kinetochore-Fiber
Ultrastructure Using
Correlative Light-Electron
Microscopy
Daniel G. Booth
*
,{,1
, Liam P. Cheeseman
*
,1
, Ian A. Prior
*
and Stephen J. Royle
*
,
{
*
Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University
of Liverpool, Liverpool, United Kingdom
{
Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, United Kingdom
{
Centre for Mechanochemical Cell Biology, Division of Biomedical Cell Biology, Warwick Medical
School, University of Warwick, Coventry, United Kingdom
CHAPTER OUTLINE
Introduction ............................................................................................................ 328
20.1 Materials........................................................................................................329
20.2 Methods .........................................................................................................331
20.2.1 Cell Transfection and Observation ................................................ 331
20.2.2 Fixation and Sample Preparation ................................................. 331
20.2.2.1 Fixative Solution Osmolarity................................................ 331
20.2.2.2 Light Microscopy ............................................................... 332
20.2.2.3 Resin Embedding .............................................................. 333
20.2.3 Longitudinal Sectioning .............................................................. 333
20.2.4 Orthogonal Sectioning................................................................. 336
20.2.5 Imaging and Sample Tilting......................................................... 336
20.3 Discussion......................................................................................................338
Acknowledgments ................................................................................................... 341
References ............................................................................................................. 341
1
These authors contributed equally to this work.
