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@PHDTHESIS{Ledesch:851775,
      author       = {Ledesch, Ralph},
      title        = {{S}imultaneous dual-color imaging on single-molecule level
                      on a {W}idefield microscope and applications},
      volume       = {178},
      school       = {RWTH Aachen},
      type         = {Dr.},
      address      = {Jülich},
      publisher    = {Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag},
      reportid     = {FZJ-2018-05290},
      isbn         = {978-3-95806-348-8},
      series       = {Schriften des Forschungszentrums Jülich. Reihe
                      Schlüsseltechnologien / Key Technologies},
      pages        = {IX, 119 S.},
      year         = {2018},
      note         = {RWTH Aachen, Diss., 2018},
      abstract     = {The epifluorescence microscope, as we know it today, has
                      come a long way. Starting as a spin-off to the UV microscope
                      at the beginning of the twentieth century, its development
                      over the last century has made it a powerful tool, allowing
                      the study of biological processes on single-molecule level
                      in unprecedented detail. The first fluorescence microscope,
                      with UV illumination, was developed at Carl Zeiss by the
                      german physicists Otto Heimstaedt and Heinrich Lehmann.
                      Between 1925 and 1932, Philipp Ellinger and August Hirt from
                      Heidelberg conceived the UV intravital microscope, that
                      allowed them to study the distribution of previously
                      injected fluorescent dyes in living kidney and liver of
                      frogs and mice [5, 6] to study their deposition and
                      transport through the blood vessels. The setup is considered
                      the prototype epifluorescence microscope: Unlike the
                      previously built transmitted light microscopes, where the
                      illumination light source is transmitted from the opposite
                      side of the specimen from the objective, in their setup, the
                      objective itself acted as the illumination condenser. While
                      the emitted red-shifted fluorescence was transmitted and
                      imaged on a diapositive, the reflected UV light was blocked
                      by a yellow filter, placed between the objective and the
                      ocular. In contrast to the transmitted light microscope, the
                      illumination light is not detected, resulting in a higher
                      image contrast, while at the same time alignment problems
                      could be avoided. The evolution to the modern
                      epifluorescence microsope is due to the contributions of the
                      russian scientist Evgenii Brumberg (State Optical Institue
                      of Leningrad) and Johann Sebastiaan Ploem, a microscopist
                      from the University of Amsterdam. They are responsible for
                      the development of the dichroic beamsplitter, a key element
                      in every modern fluorescence microscope. It physically
                      separated the excitation light from the much weaker
                      fluorescence signal, by deflecting the unwanted
                      back-reflected excitation light [7]. Up to the 1950s,
                      fluorescence microscopes used excitation light ranging from
                      the UV to the blue spectra, that was isolated from (mercury
                      or xenon) arc lamps by optical filters. The steadily
                      increasing number of developed fluorophores were not
                      necessarily optimally illuminated within the UV spectra
                      only. In 1962, Ploem collaborated with the Schott glass
                      company in Mainz to extend the spectral range for
                      illumination, by producing dichroic filters (or mirrors),
                      that deflected the blue and green spectra [8]. Furthermore,
                      Ploem collaborated with the Ernst Leitz company, which
                      constructed the first inverted microscope with
                      epi-illumination and combined the optical filter(s) and the
                      dichroic mirror in a unit (the filter cube). The filter
                      cubes were arranged to match the excitation/emission spectra
                      of the employed fluorophore. Mounted in a turret below the
                      objective, they could be interchanged [...]},
      cin          = {ICS-5},
      cid          = {I:(DE-Juel1)ICS-5-20110106},
      pnm          = {899 - ohne Topic (POF3-899)},
      pid          = {G:(DE-HGF)POF3-899},
      typ          = {PUB:(DE-HGF)3 / PUB:(DE-HGF)11},
      urn          = {urn:nbn:de:0001-2018091935},
      url          = {https://juser.fz-juelich.de/record/851775},
}