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Chapters
- 00:04Title
- 00:54Physical Alignment of Image Splitter Generated Images
- 02:25Alignment of Spectrally Separated Channels by Software Performing Spatial Transformation
- 04:34Single Molecule Imaging of Mitochondrial Membrane Proteins
- 08:32Results: Multi-color Localization of Mammalian Mitochondrial PINK1
- 10:15Conclusion
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.
Tags
Keywords: Multi-color Localization MicroscopySingle Membrane ProteinsOrganellesLive Mammalian CellsProtein DynamicsSpatiotemporal OrganizationMitochondrial Membrane ProteinsTransfectionSample PreparationMicroscope SetupFluorescent BeadsObjective LensImmersion OilTransmission LightLaserFluorescence ChannelsImage SplitterTIRF MicroscopyTransformation MatrixDual-color ImagesCalibrationUnit ConversionLocalization ManagerPoint Spread Function (PSF)
