title: Fluorescence live cell imaging.
abstract: Fluorescence microscopy of live cells has become an integral part of modern cell biology. Fluorescent protein (FP) tags, live cell dyes, and other methods to fluorescently label proteins of interest provide a range of tools to investigate virtually any cellular process under the microscope. The two main experimental challenges in collecting meaningful live cell microscopy data are to minimize photodamage while retaining a useful signal-to-noise ratio and to provide a suitable environment for cells or tissues to replicate physiological cell dynamics. This chapter aims to give a general overview on microscope design choices critical for fluorescence live cell imaging that apply to most fluorescence microscopy modalities and on environmental control with a focus on mammalian tissue culture cells. In addition, we provide guidance on how to design and evaluate FP constructs by spinning disk confocal microscopy.
authors: Department of Cell and Tissue Biology, University of California, San Francisco, USA.
authors: Andreas
authors: A
authors: Ettinger
authors: Department of Cell and Tissue Biology, University of California, San Francisco, USA.
authors: Torsten
authors: T
authors: Wittmann
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D038761:Photobleaching; D059629:Signal-To-Noise Ratio; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D049452:Green Fluorescent Proteins
keywords: Environmental control; Fluorescence microscopy; Fluorescent proteins; Live cell microscopy; Photobleaching; Spinning disk confocal microscopy