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Journal / Bioengineering / Photoactivated Localization Microscopy with…
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JoVE Journal Bioengineering
Photoactivated Localization Microscopy with Bimolecular Fluorescence Complementation (BiFC-PALM)
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Photoactivated Localization Microscopy with Bimolecular Fluorescence Complementation (BiFC-PALM)

Article DOI: 10.3791/53154-v • 12:42 min • December 22nd, 2015
December 22nd, 2015 •
1,2,3, 1,2,3, 1,2,3, 1,2,3
1Department of Biomedical Engineering, Oregon Health and Science University, 2Knight Cancer Institute, Oregon Health and Science University, 3OHSU Center for Spatial Systems Biomedicine, Oregon Health and Science University

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Summary
Automatic Translation

Protein-protein interactions are visualized in cells with nanometer spatial resolution by combining bimolecular fluorescence complementation (BiFC) with photoactivated localization microscopy (PALM). Described here is the use of BiFC-PALM for imaging Ras-Raf interactions in U2OS cells for visualizing the nanoscale clustering and diffusion of individual Ras-Raf complexes.

Tags

Photoactivated Localization Microscopy Bimolecular Fluorescence Complementation Protein-protein Interactions Visualization Resolution Sensitivity Cells Conventional Light Microscopy Diffraction-limited Bimolecular Fluorescence Complementation Photoactivated Localization Microscopy Single Molecule Sensitivity Nanometer Spatial Resolution Fluorescent Protein PALM Technique Superresolution Microscopy Phototransformable Fluorescent Probes Photoactivatable Fluorescent Proteins PAmCherry1 Monomeric Nature Single Molecule Brightness Contrast Ratio Stoichiometry Measurements BiFC Probe Amino Acids 159 And 160 High Specificity Non-specific Reconstitution
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