Fluorescence live cell imaging.
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.
['Ettinger|Andreas|A|', 'Wittmann|Torsten|T|']
D000818:Animals D002478:Cells, Cultured D049452:Green Fluorescent Proteins D006801:Humans D018613:Microscopy, Confocal D008856:Microscopy, Fluorescence D038761:Photobleaching D059629:Signal-To-Noise Ratio D059010:Single-Cell Analysis
Fluorescence microscopy Fluorescent proteins Live cell microscopy Environmental control Spinning disk confocal microscopy Photobleaching