3D-STED Microscope

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Investigations of small subcellular compartments has been significantly improved by the use of localisation microscopy methods. These methods allow to look into the structure and dynamic of molecular signalling complexes and are today standard microscopy tools in cell biology. The gain of localisation precision is about tenfold in comparison to confocal microscopy and reach the resolution of electron microscopy. The main advantage is the much simpler preparation of the probes and the possibility to use localisation microscopy also in living cells. Stochastic approach as Photoactivation localisation microscopy (PALM) or Stochastic Optical Reconstruction Microscopy (STORM) is most obvious in the gain of single fluorophore information, but relay on the acquisition of many pictures and precise reconstructions. The deterministic approach, called Stimulated Emission Depletion microscopy (STED) use a defined light pattern to scan the sample and allow to correlate much faster the localisation of molecules to each other. The optimization of detectors and illumination algorithms has improved the usability of STED microscopy in living cells. Implementation of adaptive optics or corrections in the optical components allow also a nearly similar localisation precision in all three spatial dimensions. With these improvements the STED microscopy is now relevant for a broad spectrum of samples. With the acquisition of a STED microscope we like to provide a valid tool for the growing number of molecular/cell biological working groups in the biology department of the Johannes Gutenberg University Mainz to conduct their research. The use of pulsed lasers within the STED microscope offers in addition the possibility to use the fluorescent live time of the fluorophores and investigate their interactions in relevant dimensions to their local arrangement.