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.
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Acknowledgements
This work was supported by Invitation Fellowship Programs for Research in Japan (no. L-08551) to T.B. and Grant-in-Aid for Scientific Research (B) (no. 21370026) to T.M. from the Japanese Society for Promotion of Science.
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Murata, T., Baskin, T.I. (2014). Imaging the Mitotic Spindle by Spinning Disk Microscopy in Tobacco Suspension Cultured Cells. In: Sharp, D. (eds) Mitosis. Methods in Molecular Biology, vol 1136. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0329-0_4
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DOI: https://doi.org/10.1007/978-1-4939-0329-0_4
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