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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
 

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Article DOI: 10.3791/57690-v • 11:06 min • June 30th, 2018
June 30th, 2018 •
1, 1, 1, 2, 1,3
1School of Biology, University of Osnabrück, 2Center of Cellular Nanoanalytics, Integrated Bioimaging Facility, University of Osnabrück, 3Department of Biology, WWU Münster

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Here, we present a protocol for multi-color localization of single membrane proteins in organelles of live cells. To attach fluorophores, self-labeling proteins are used. Proteins, located in different membranes compartments of the same organelle, can be localized with a precision of ~18 nm.

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Keywords: Multi-color Localization Microscopy Single Membrane Proteins Organelles Live Mammalian Cells Protein Dynamics Spatiotemporal Organization Mitochondrial Membrane Proteins Transfection Sample Preparation Microscope Setup Fluorescent Beads Objective Lens Immersion Oil Transmission Light Laser Fluorescence Channels Image Splitter TIRF Microscopy Transformation Matrix Dual-color Images Calibration Unit Conversion Localization Manager Point Spread Function (PSF)
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