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The Stedycon unit was installed on a Zeiss Axiovert OBSERVER Z1 microscope enabling for live cell imaging, widefield epifluorescence, confocal and STED microscopy. A piezo-driven 100x objective allows 3D acquisition. An electronic YX-stage is controlled by joystick. A halogen lamp and an HBO lamp give widefield illumination for brightfield and fluorescence images, respectively.
Univ.-Prof. Dr. Silja Weßler
STED Mikroskopie, Live Cell Imaging
Methods & Expertise for Research Infrastructure
The Sted micrsocope will be used for different molecular biological and cell biological research. The StedyCon allows confocal and super-resolution microscopyand allows super-resolved analysis of fixed and living cells. Superresolution microscopy using stimulated emission depletion (STED) creates sub-diffraction limit features by altering the effective point spread function of the excitation beam using a second laser that suppresses fluorescence emission from fluorophores located away from the center of excitation. The suppression of fluorescence is achieved through stimulated emission that occurs when an excited-state fluorophore encounters a photon that matches the energy difference between the ground and excited state. Upon interaction of the photon and the excited fluorophore, the molecule is returned to the ground state through stimulated emission before spontaneous fluorescence emission can occur. Thus, the process effectively depletes selected regions near the focal point of excited fluorophores that are capable of emitting fluorescence.