Journal Article

Light Sheet Microscopy for Single Molecule Tracking in Living Tissue

Ritter, Jörg Gerhard, Veenendaal, Andreas, Veith, Roman, Siebrasse, Jan Peter, Kubitscheck, Ulrich

2010

Overview

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 microm 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.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Animals / Biomolecules / Biophysics / Biophysics/Experimental Biophysical Methods / Cell Nucleus - metabolism / Chironomidae - metabolism / Chironomus tentans / Experiments / Feasibility studies / Fluorescence / Fluorescence microscopy / Gene expression / Larvae / Light / Localization / Microscopy / Microscopy, Fluorescence - methods / Molecular Biology/mRNA Transport and Localization / Molecular diffusion / Nuclei / Nuclei (cytology) / Optical sectioning / Physiological aspects / Proteins / Ribonucleoproteins - metabolism / Salivary gland / Salivary Glands - metabolism / Sectioning / Signal quality / Tracking