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