title: Best practices for fluorescence microscopy of the cyanobacterial circadian clock.
abstract: This chapter deals with methods of monitoring the subcellular localization of proteins in single cells in the circadian model system Synechococcus elongatus PCC 7942. While genetic, biochemical, and structural insights into the cyanobacterial circadian oscillator have flourished, difficulties in achieving informative subcellular imaging in cyanobacterial cells have delayed progress of the cell biology aspects of the clock. Here, we describe best practices for using fluorescent protein tags to monitor localization. Specifically, we address how to vet fusion proteins and overcome challenges in microscopic imaging of very small autofluorescent cells.
authors: Center for Circadian Biology, University of California, San Diego, La Jolla, California, USA; Division of Biological Sciences, University of California, San Diego, La Jolla, California, USA.
authors: Susan E
authors: SE
authors: Cohen
authors: Division of Biological Sciences, University of California, San Diego, La Jolla, California, USA.
authors: Marcella L
authors: ML
authors: Erb
authors: Division of Biological Sciences, University of California, San Diego, La Jolla, California, USA.
authors: Joe
authors: J
authors: Pogliano
authors: Center for Circadian Biology, University of California, San Diego, La Jolla, California, USA; Division of Biological Sciences, University of California, San Diego, La Jolla, California, USA. Electronic address: sgolden@ucsd.edu.
authors: Susan S
authors: SS
authors: Golden
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D057906:Circadian Clocks; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial / Q000528:radiation effects; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008027:Light; D008856:Microscopy, Fluorescence; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D046940:Synechococcus / Q000502:physiology*; D059008:Time-Lapse Imaging
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins
keywords: Cyanobacteria; Fluorescence imaging; GFP; Subcellular localization