title: Ultramicroscopy: development and outlook.
abstract: We present an overview of the ultramicroscopy technique we developed. Starting from developments 100 years ago, we designed a light sheet microscope and a chemical clearing to image complete mouse brains. Fluorescence of green fluorescent protein (GFP)-labeled neurons in mouse brains could be preserved with our 3DISCO clearing and high-resolution three-dimensional (3-D) recordings were obtained. Ultramicroscopy was also used to image whole mouse embryos and flies. We improved the optical sectioning of our light sheet microscope by generating longer and thinner light sheets with aspheric optics. To obtain high-resolution images, we corrected available air microscope objectives for clearing solutions with high refractive index. We discuss how eventually super resolution could be realized in light sheet microscopy by applying stimulated emission depletion technology. Also the imaging of brain function by recording of mouse brains expressing cfos-GFP is discussed. Finally, we show the first 3-D recordings of human breast cancer with light sheet microscopy as application in medical diagnostics.
authors.affiliation: Vienna University of Technology, FKE, Chair of Bioelectronics, Floragasse 7, 1040 Vienna, Austria; Medical University of Vienna, Center for Brain Research, Spitalgasse 4, 1090 Vienna, Austria.
authors.forename: Hans-Ulrich
authors.initials: HU
authors.lastname: Dodt
authors.affiliation: Vienna University of Technology, FKE, Chair of Bioelectronics, Floragasse 7, 1040 Vienna, Austria; Medical University of Vienna, Center for Brain Research, Spitalgasse 4, 1090 Vienna, Austria.
authors.forename: Saiedeh
authors.initials: S
authors.lastname: Saghafi
authors.affiliation: Vienna University of Technology, FKE, Chair of Bioelectronics, Floragasse 7, 1040 Vienna, Austria; Medical University of Vienna, Center for Brain Research, Spitalgasse 4, 1090 Vienna, Austria.
authors.forename: Klaus
authors.initials: K
authors.lastname: Becker
authors.affiliation: Vienna University of Technology, FKE, Chair of Bioelectronics, Floragasse 7, 1040 Vienna, Austria; Medical University of Vienna, Center for Brain Research, Spitalgasse 4, 1090 Vienna, Austria.
authors.forename: Nina
authors.initials: N
authors.lastname: Jährling
authors.affiliation: Vienna University of Technology, FKE, Chair of Bioelectronics, Floragasse 7, 1040 Vienna, Austria; Medical University of Vienna, Center for Brain Research, Spitalgasse 4, 1090 Vienna, Austria.
authors.forename: Christian
authors.initials: C
authors.lastname: Hahn
authors.affiliation: Vienna University of Technology, FKE, Chair of Bioelectronics, Floragasse 7, 1040 Vienna, Austria; Medical University of Vienna, Center for Brain Research, Spitalgasse 4, 1090 Vienna, Austria.
authors.forename: Marko
authors.initials: M
authors.lastname: Pende
authors.affiliation: Vienna University of Technology, FKE, Chair of Bioelectronics, Floragasse 7, 1040 Vienna, Austria; Medical University of Vienna, Center for Brain Research, Spitalgasse 4, 1090 Vienna, Austria.
authors.forename: Martina
authors.initials: M
authors.lastname: Wanis
authors.affiliation: Pathologie Hamburg-West , Lornsenstraße 4, 22767 Hamburg, Germany.
authors.forename: Axel
authors.initials: A
authors.lastname: Niendorf
keywords: brain clearing; light sheet; objective correction; three-dimensional brain imaging; tumor clearing; ultramicroscopy