Lehrstuhl für Biotechnologie und Biophysik

    Re-scan Confocal Microscopy (RCM)

    We set up a correlative system using the super-resolution technique dSTORM and Re-scan Confocal Microscopy (RCM), which is a resolution-enhanced form of LSM. By re-scanning the emission light after the pinhole, RCM gains a resolution improvement of 1.4 fold. This will increase the lateral resolution from 240 nm of a confocal microscope to 170 nm of the RCM, while maintaining the sectioning capability of a confocal system. Since the detector (Photomultiplier Tube) is replaced by a sCMOS camera with high quantum efficiency, the signal-to-noise ratio of the RCM is twice as good as for a confocal microscope. With this system we are now capable of quantitatively investigating protein distributions in the context of dynamic processes with high spatial resolution.

    Reference: de Luca et al., 2013

    Microscope-body: Nikon TiE with motorized stage and PFS-System (Perfect-Focus-System), inverted
    Objective: 100x Oil, NA 1.49 Apo-TIRF
    60x Water, NA 1.27
    100x Siliconoil, 1.3
    dSTORM Detection
    Laser: 405nm, 488nm, 640nm: 170mW, 532nm: 500mW
    Camera:EMCCD, Andor iXON DU-897
    Laser: Cobolt Skyra, Multiline lasersystem
    405nm, 488nm, 561nm, 640nm: 50mW
    Camera:sCMOS, Andor Zyla 4.2P
    Timelapse of Hela cells expressing EB1 (end binding protein of tubulin) tagged with YFP
    COS7 cell with staining of nucleus (Hoechst), actin (Phalloidin 488) and microtubuli (Alexa Fluor 647)

    Lehrstuhl für Biotechnologie und Biophysik
    Am Hubland
    97074 Würzburg

    Tel.: +49 931 31-84507

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    Hubland Süd, Geb. B1 Hubland Nord, Geb. 32 Julius-von-Sachs-Platz 2 Fabrikschleichach Hubland Süd, Geb. B2 Campus Medizin