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Oligomerization Dynamics of Cell Surface Receptors in Living Cells by Total Internal Reflection Fluorescence Microscopy Combined with Number and Brightness Analysis

Article DOI: 10.3791/60398-v • 10:43 min • November 6th, 2019
November 6th, 2019 •
1, 2, 1, 3, 1,2
1Unit of Microscopy and Dynamic Imaging, Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain, 2Centro di Imaging Sperimentale, Ospedale San Raffaele, Milan, Italy, 3Unit of Gynecological Oncology Research, European Institute of Oncology IRCCS, Milan, Italy

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Summary
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We describe an imaging approach for the determination of the average oligomeric state of mEGFP-tagged-receptor oligomers induced by ligand binding in the plasma membrane of living cells. The protocol is based on Total Internal Reflection Fluorescence (TIRF) microscopy combined with Number and Brightness (N&B) analysis.

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Oligomerization Dynamics Cell Surface Receptors Living Cells Total Internal Reflection Fluorescence Microscopy Number And Brightness Analysis Receptor Clustering TIRF Microscopy FGFR1-mEGFP Molecules Plasma Membrane Stimulated Receptor Clustering Spatial Temporal Organization Fluorescence Fluctuation Methods Stoichiometrically Labeled Intracellular Compartments Monomeric Form Of Fluorophore
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