Max Delbruck Center for Molecular Medicine 2 articles published in JoVE 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 Friedrich1, Johannes Balkenhol1, Felix Knote1, Martin J. Lohse2, Katrin G. Heinze1 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. Biology Isolation, Culture, and Genetic Engineering of Mammalian Primary Pigment Epithelial Cells for Non-Viral Gene Therapy Thais Bascuas*1,2, Martina Kropp*1,2, Nina Harmening1,2, Brittany M. Wong1, Sandra Johnen3, Zsuzsanna Izsvák4, Gabriele Thumann1,2 1Experimental Ophthalmology, University of Geneva, 2Department of Ophthalmology, University Hospitals of Geneva, 3Department of Ophthalmology, University Hospital RWTH Aachen, 4Max Delbrück Center for Molecular Medicine Here, a protocol to isolate and transfect primary iris and retinal pigment epithelial cells from various mammals (mice, rat, rabbit, pig, and bovine) is presented. The method is ideally suited to study ocular gene therapy approaches in various set-ups for ex vivo analyses and in vivo studies transferable to humans.