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JoVE Journal Bioengineering
High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy
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High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

Article DOI: 10.3791/51784-v • 15:13 min • July 25th, 2014
July 25th, 2014 •
1, 1, 1, 1
1Institute of Pharmacology and Toxicology and Bio-Imaging Center/Rudolf Virchow Center, DFG-Research Center for Experimental Biomedicine, University of Würzburg, Germany

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Summary
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This protocol describes how to use total internal reflection fluorescence microscopy to visualize and track single receptors on the surface of living cells and thereby analyze receptor lateral mobility, size of receptor complexes as well as to visualize transient receptor-receptor interactions. This protocol can be extended to other membrane proteins.

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High-resolution Spatiotemporal Analysis Receptor Dynamics Single-molecule Fluorescence Microscopy Signaling Molecules Ensemble Measurements Biochemical Methods Microscopy Methods Dynamic Events Receptor-receptor Interactions Living Cell Spatiotemporal Resolution Labeling With Organic Fluorophores Total Internal Reflection Fluorescence Microscopy (TIRF Microscopy) Visualizing Single Receptors Sample Preparation Image Acquisition Image Analysis G-protein-coupled Receptors β2-adrenergic Receptor γ-aminobutyric Acid Type B (GABAB) Receptor Membrane Proteins Cell Models Transfection Methods Labeling Strategies
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