title: Imaging the mitotic spindle by spinning disk microscopy in tobacco suspension cultured cells.
abstract: Plants are valuable systems for analyzing the acentriolar mitotic spindle. We have developed methods for imaging the mitotic spindle in living tobacco (Nicotiana tabacum) suspension culture cells expressing GFP-α-tubulin. The methods allow the spindle to be observed in living cells at high spatial and temporal resolution and rely on water immersion objectives, spinning disk optics, and high-sensitivity cameras. Here, we describe these methods and provide step-by-step protocols for certain key steps. We also describe a method for application and removal of inhibitors.
authors.affiliation: Department of Basic Biology, School of Life Science, The Graduate University for Advanced Studies Nishigonaka, 38 Myodaiji-cho, Okazaki, Aichi, 444-8585, Japan, tkmurata@nibb.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Murata
authors.forename: Tobias I
authors.initials: TI
authors.lastname: Baskin
mesh_terms: D002478:Cells, Cultured; D020090:Chromosome Segregation; D015870:Gene Expression; D017930:Genes, Reporter; D018613:Microscopy, Confocal / Q000379:methods; D008870:Microtubules / Q000378:metabolism; D059828:Plant Cells; D008941:Spindle Apparatus / Q000378:metabolism*; D059008:Time-Lapse Imaging; D014026:Nicotiana / Q000166:cytology*