Single marker switching nanoscopy with 4Pi detection [Elektronische Ressource] : superior nanometric 3D resolution / presented by Daniel Aquino Maier

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-Physiker Daniel Aquino Maierborn in CochabambaOral examination: February 1st, 2011Single Marker Switching Nanoscopywith 4Pi Detection:Superior Nanometric 3D ResolutionReferees: Prof. Dr. Dr. h.c. Stefan W. HellProf. Dr. Rainer H. A. FinkvAbstract – ZusammenfassungThe spatial resolution of light microscopy was limited by diffraction for a long time. Switch-ing markers between a dark and a bright state was the key element to overcome this limitation.In single marker switching (SMS) microscopy, switching and subsequent localization of singlemarkers enables the creation of two-dimensional superresolution images. The approaches to ex-tend SMS microscopy to the third dimension were hitherto hampered by a substantially anisotropicresolution or limited to samples thinner than half the marker fluorescence emission wavelength.Still, a light microscope allowing three-dimensional subdiffraction imaging of thick samples witha homogeneous and isotropic resolution combined with the ability to differentiate between differ-ent types of markers is highly desirable.This thesis presents a nanoscope which combines the SMS concept with the 4Pi-technique to ad-dress these important issues.