Light Sheet Microscopy to Measure Protein Dynamics.
Visualizing protein dynamics is the key to a quantitative understanding of molecular mechanisms in biological systems. Recent developments in fluorescent microscopy techniques allow for novel experimental approaches to study stochastic localization, distribution, and movement of fluorescently labeled molecules in high resolution as they occur over time in the context of living cells and whole organisms. Particularly suitable for such studies is the application of light sheet microscopy, as it enables rapid in vivo imaging of a wide range and size of specimen, from whole organisms and organs, down to the dynamics of subcellular structures and single molecules. This article summarizes the principles of light sheet microscopy and its advantages over other optical imaging techniques, such as light microscopy and confocal microscopy, and highlights recent innovations that significantly enhance spatio-temporal resolution. Also, this manuscript contains basic guidelines for the implementation of light sheet microscopy in the laboratory and presents thus far unpublished light sheet microscopy applications demonstrating the ability of this technology to measure protein dynamics in a whole-organism context. J. Cell. Physiol. 232: 27-35, 2017. © 2016 Wiley Periodicals, Inc.
Institute for Genome Stability in Ageing and Disease, Cologne Cluster of Excellence in Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany. m_rieckher@gmx.com, mrieckh1@uni-koeln.de.
Matthias
M
Rieckher
D000818:Animals; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008027:Light; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence / Q000379:methods; D061848:Optical Imaging / Q000295:instrumentation / Q000379:methods; D021381:Protein Transport; D011506:Proteins / Q000378:metabolism*
D011506:Proteins