The Nikon N-STORM microscope at the MRC Laboratory for Molecular Cell Biology Super-Resolution Facility (SuRF) uses STochastic Optical Reconstruction Microscopy (STORM) localization with which lateral resolutions of ~20 nm in biological specimens can be achieved. This allows the study of protein-protein interactions at the molecular level using localization information of individual fluorophores within the specimen. Axial resolutions of ~50 nm can be reached using a cylindrical lens that induces a known astigmatic stretch to the image therefore allowing super-resolution information to be obtained in 3D. The system is built around the Nikon Ti-E inverted microscope and allows high-accuracy, multi-colour localization. It is equipped with an Lu-NV laser bed providing four laser lines (405, 488, 561 and 647 nm) for sample excitation, an Andor iXon EM-CCD camera for fluorescence detection and comes with the NIS-Elements software package for image acquisition and processing. The 100x oil TIRF CFI Apochromat (NA 1.49) objective lens is ideal for STORM imaging and incubation equipment for live cell experiments is also available. Multi-colour STORM imaging can be performed using tandem dye pairs or standard secondary antibodies in continuous activation mode. The microscope is located in room G-08.
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