Expanding the STED principle to multicolor imaging and far-field optical writing [Elektronische Ressource] / presented by Lars Meyer

INAUGURAL - DISSERTATIONsubmitted to theCombined Faculties for the Natural Sciences and for Mathematicsof the Ruperto - Carola University of Heidelberg, Germanyfor the degree ofDoctor of Natural Sciencespresented byDiplom-Chemiker Lars Meyerborn in VechtaOral examination: December 22, 2009Expanding the STED PrincipletoMulticolor Imaging and Far-Field Optical WritingReferees: Prof. Dr. Joachim SpatzProf. Dr. Stefan W. HellAbstractStimulated emission depletion (STED) nanoscopy has emerged as a powerful far-field tech-nique for subdiffraction optical imaging and is extensively used in the life sciences to in-vestigate different protein species. In many cases, however, the relative assembly of two (ormore) proteins is of interest and needs to be determined with nanometric resolution. Mean-while, STED has also found its way into material science. Yet, the method’s potential forsubdiffraction optical writing has so far remained unexplored. This work evidences the de-velopment of a dual-color STED setup with lateral resolving power of 30 nm in both colorchannels. The method is shown to be applicable to the study of double-stained neuronal pro-teins and aids in establishing a novel sample sectioning technique. As a result the first high-resolution three-dimensional reconstruction of dual-color STED images is rendered possible.Flexible operation of the individual color channels furthermore answers open questions inchemistry and biology.