Parallelized ground state depletion [Elektronische Ressource] / by Miriam A. Schwentker

DissertationsubmittedtotheCombinedFacultyfor theNaturalSciencesandforMathmaticsoftheRuperto CarolaUniversityofHeidelberg,GermanyforthedegreeofDoctorofNaturalSciencesbyDiplom PhysikerinMiriamA.Schwentkerborn13.07.1977inAugsburgoralexam: 06.06.2007GelängeunsdochderGriffausdemDunkelnachdenwirklichenDingen!(HeinzCzechowski)ParallelizedGroundStateDepletionGutachter: Prof. Dr. StefanW.HellProf. Dr. JosefBilleAbstractRESOLFT was introduced to break the limited resolution in fluorescence microscopygiven by thephysical limit of diffraction. RESOLFT needs saturable deple tion and illumination with spatial regions of zero intensity. Improvements in RESOLFTare still essential since (i) photobleaching restricts the use of large intensities obligate forhigh resolution, (ii) long acquisition times emerge from single point scanning, and (iii) RE SOLFTputsaheadspecialrequirementsonthedye´sphotophysicalproperties. Tocounteractthese problems, this work presents a RESOLFT type microscope implementing parallelizedground state depletion (GSD). Parallelization allows rather fast image acquisition apply ing structured illumination combined with widefield detection on a camera. The depletionof the fluophore´s ground state, and thus the fluorescence, is realized using a pump probeilluminationscheme. Toincreasephotostability,thesamplewasevacuatedandcooledtoap proximately80K.