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FRENCH INFRASTRUCTURE FOR INTEGRATED STRUCTURAL BIOLOGY

Super-resolution-localization-microscopy.jpg

INSTRUCT CENTER - FRANCE 1 - IGBMC

SUPER RESOLUTION LOCALIZATION MICROSCOPY

The SR GSD allows a range of localization super-resolution experiments (dSTORM, GSDIM, STORM, PALM), particle tracking, standard wide-field epifluorescence and TIRF microscopy, transmission microscopy with polarization contrast. Samples can be mounted either on standard slides or in 35mm diameter glass bottom petri dishes.

The Leica SR GSD system is based on the Leica DMI6000 B inverted microscope that equipped with a TIRF attachment, 100x/1.47 oil immersion and 10x/0.40 dry objectives. The especially for GSDIM designed SuMo sample stage with objective linked to the stage and active anti-vibration table ensures low sample drift (rated of <20 nm in 10 min). The maximal achievable resolution, depending on specimen, lies in range from 20 to 70 nm; the localization precision of individual fluorophores can be as high as a few nanometers.

Different laser lines can be used for excitation of fluorescence and photoconversion: 488 nm 300 mW, 532 nm 1000 mW, 642 nm 500 mW, 405 nm 50 mW. The images are captured by the Andor iXon3 897 EMCCD camera with equivalent pixel size of 100 nm. The field of view with the 100x objective and a 1.6x magnification lens is 51x51 µm in conventional wide-field mode and 18x18 µm during super-resolution imaging.

The microscope is additionally equipped with the MicAO 3D-SR adaptive optics system that permits detection and correction of optical aberrations in the sample as well as creation of a perfect astigmatism for 3D experiments.

The microscope is driven by the LAS AF software that allows full control of experiments and super-resolution image reconstruction, in real time as well as after the acquisition with various post-processing options.

The microscopy room is equipped with basic tools allowing mounting of samples and their storage at +4 °C/-20 °C.

Device: Leica SR GSD