Imaging Polarization in Budding Yeast.
We describe methods for live-cell imaging of yeast cells that we have exploited to image yeast polarity establishment. As a rare event occurring on a fast time-scale, imaging polarization involves a trade-off between spatiotemporal resolution and long-term imaging without excessive phototoxicity. By synchronizing cells in a way that increases resistance to photodamage, we discovered unexpected aspects of polarization including transient intermediates with more than one polarity cluster, oscillatory clustering of polarity factors, and mobile "wandering" polarity sites.
Department of Pharmacology and Cancer Biology, Duke University School of Medicine, C359 LSRC, Box 3813, Durham, NC, 27710, USA.
Allison W
AW
McClure
Department of Pharmacology and Cancer Biology, Duke University School of Medicine, C359 LSRC, Box 3813, Durham, NC, 27710, USA.
Chi-Fang
CF
Wu
Light Microscopy Core Facility, Duke University, Durham, NC, USA.
Sam A
SA
Johnson
Department of Pharmacology and Cancer Biology, Duke University School of Medicine, C359 LSRC, Box 3813, Durham, NC, 27710, USA. Daniel.lew@duke.edu.
Daniel J
DJ
Lew
D016764:Cell Polarity; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D016192:Resting Phase, Cell Cycle / Q000235:genetics; D004718:Saccharomycetales / Q000166:cytology* / Q000502:physiology
D005656:Fungal Proteins
Cdc42; Cell polarity; GFP; Imaging; Microscopy; Polarity establishment; Yeast