Max Delbrück Center for Molecular Medicine, Berlin
2 ARTICLES PUBLISHED IN JoVE
Bioengineering
High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy
Titiwat Sungkaworn 1, Finn Rieken 1, Martin J. Lohse 1, Davide Calebiro 1
1Institute of Pharmacology and Toxicology and Bio-Imaging Center/Rudolf Virchow Center, DFG-Research Center for Experimental Biomedicine, University of Würzburg, Germany
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.
Biochemistry
Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
Katherina Hemmen *1, Susobhan Choudhury *1, Mike Friedrich 1, Johannes Balkenhol 1, Felix Knote 1, Martin J. Lohse 2, Katrin G. Heinze 1
1Rudolf Virchow Center for Integrative and Translation Bioimaging, Julius-Maximilian University Wuerzburg, 2Max Delbrück Center for Molecular Medicine, Berlin
We present an experimental protocol and data analysis workflow to perform live cell dual-color fluorescence cross correlation spectroscopy (FCCS) combined with Förster Resonance Energy transfer (FRET) to study membrane receptor dynamics in live cells using modern fluorescence labeling techniques.
