title: Single yeast cell imaging.
abstract: Microscopic imaging techniques play a pivotal role in the life sciences. Here we describe labeling and imaging methods for live yeast cell imaging. Yeast is an excellent reference organism for biomedical research to investigate fundamental cellular processes, and has gained great popularity also for large-scale imaging-based screens. Methods are described to label live yeast cells with organelle-specific fluorescent dyes or GFP-tagged proteins, and how cells are maintained viable over extended periods of time during microscopy. We point out common pitfalls and potential microscopy artifacts arising from inhomogeneous labeling and depending on cellular physiology. Application and limitation of bleaching techniques to address dynamic processes in the yeast cell are described.
authors.affiliation: Institute of Molecular Biosciences, University of Graz, Graz, Austria.
authors.forename: Heimo
authors.initials: H
authors.lastname: Wolinski
authors.forename: Sepp D
authors.initials: SD
authors.lastname: Kohlwein
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D018914:Cells, Immobilized / Q000166:cytology / Q000648:ultrastructure; D036681:Fluorescence Recovery After Photobleaching / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis*; D049452:Green Fluorescent Proteins / Q000032:analysis*; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D061848:Optical Imaging / Q000379:methods*; D038761:Photobleaching; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000648:ultrastructure; D013194:Staining and Labeling / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins