title: 4D Confocal Imaging of Yeast Organelles.
abstract: Yeast cells are well suited to visualizing organelles by 4D confocal microscopy. Typically, one or more cellular compartments are labeled with a fluorescent protein or dye, and a stack of confocal sections spanning the entire cell volume is captured every few seconds. Under appropriate conditions, organelle dynamics can be observed for many minutes with only limited photobleaching. Images are captured at a relatively low signal-to-noise ratio and are subsequently processed to generate movies that can be analyzed and quantified. Here, we describe methods for acquiring and processing 4D data using conventional scanning confocal microscopy.
authors.affiliation: Department of Molecular Genetics and Cell Biology, University of Chicago, 920 East 58th St., Chicago, IL, 60637, USA.
authors.forename: Kasey J
authors.initials: KJ
authors.lastname: Day
authors.affiliation: Department of Molecular Genetics and Cell Biology, University of Chicago, 920 East 58th St., Chicago, IL, 60637, USA.
authors.forename: Effrosyni
authors.initials: E
authors.lastname: Papanikou
authors.affiliation: Department of Molecular Genetics and Cell Biology, University of Chicago, 920 East 58th St., Chicago, IL, 60637, USA. bsglick@uchicago.edu.
authors.forename: Benjamin S
authors.initials: BS
authors.lastname: Glick
mesh_terms: D007091:Image Processing, Computer-Assisted / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods; D015388:Organelles; D012441:Saccharomyces cerevisiae / Q000166:cytology*
keywords: 4D microscopy; Confocal; Deconvolution; ImageJ; Photobleaching; Yeast