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Light Sheet Microscopy for Single Molecule Tracking in Living Tissue

Authors
Publisher
Public Library of Science
Publication Date
Volume
5
Issue
7
Identifiers
DOI: 10.1371/journal.pone.0011639
Keywords
  • Research Article
  • Biophysics
  • Biophysics/Experimental Biophysical Methods
  • Molecular Biology/Mrna Transport And Localization
Disciplines
  • Biology

Abstract

Single molecule observation in cells and tissue allows the analysis of physiological processes with molecular detail, but it still represents a major methodological challenge. Here we introduce a microscopic technique that combines light sheet optical sectioning microscopy and ultra sensitive high-speed imaging. By this approach it is possible to observe single fluorescent biomolecules in solution, living cells and even tissue with an unprecedented speed and signal-to-noise ratio deep within the sample. Thereby we could directly observe and track small and large tracer molecules in aqueous solution. Furthermore, we demonstrated the feasibility to visualize the dynamics of single tracer molecules and native messenger ribonucleoprotein particles (mRNPs) in salivary gland cell nuclei of Chironomus tentans larvae up to 200 µm within the specimen with an excellent signal quality. Thus single molecule light sheet based fluorescence microscopy allows analyzing molecular diffusion and interactions in complex biological systems.

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