Abstract
The discovery of green fluorescent protein (GFP) allowed visualization of a wide variety of processes within living cells. Thanks to the development of differently colored fluorophores, it is now possible to simultaneously follow distinct subcellular events at the single cell level. Here, we describe a basic method to visualize multiple events during cytokinesis by time-lapse fluorescence microscopy in the budding yeast Saccharomyces cerevisiae. In this organism, contraction of an actomyosin-based ring drives ingression of the plasma membrane at the mother–bud division site to partition the cytoplasm of the dividing cell. Simultaneous visualization of distinct cytokinesis steps in living cells, such as ring contraction and membrane ingression, will facilitate a complete understanding of the mechanisms of eukaryotic cell division.
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Acknowledgments
We thank all lab members for helpful suggestions. Research in our laboratory is supported by the Spanish ministry of Economy and Competitiveness (BFU09-08213).The research leading to these results has received funding from the European Research Council under the European Union's Seventh Framework Programme (FP7/2007-2013) / ERC grant agreement n° 260965. It reflects only the authors’ views and the Union is not liable for any use that may be made of the information contained therein.
We acknowledge support of the Spanish Ministry of Economy and Competitiveness, ‘Centro de Excelencia Severo Ochoa 2013-2017’, SEV-2012-0208.
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Kumar, A., Mendoza, M. (2016). Time-Lapse Fluorescence Microscopy of Budding Yeast Cells. In: Sanchez-Diaz, A., Perez, P. (eds) Yeast Cytokinesis. Methods in Molecular Biology, vol 1369. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3145-3_1
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DOI: https://doi.org/10.1007/978-1-4939-3145-3_1
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3144-6
Online ISBN: 978-1-4939-3145-3
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