title: Design and performance of an ultra-flexible two-photon microscope for in vivo research.
abstract: We present a cost-effective in vivo two-photon microscope with a highly flexible frontend for in vivo research. Our design ensures fast and reproducible access to the area of interest, including rotation of imaging plane, and maximizes space for auxiliary experimental equipment in the vicinity of the animal. Mechanical flexibility is achieved with large motorized linear stages that move the objective in the X, Y, and Z directions up to 130 mm. 360° rotation of the frontend (rotational freedom for one axis) is achieved with the combination of a motorized high precision bearing and gearing. Additionally, the modular design of the frontend, based on commercially available optomechanical parts, allows straightforward updates to future scanning technologies. The design exceeds the mobility of previous movable microscope designs while maintaining high optical performance.
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Johannes M
authors.initials: JM
authors.lastname: Mayrhofer
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland ; IZKF Aachen, Medical Faculty of the RWTH Aachen University, Pauwelsstraße 30, 52074 Aachen, Germany ; Institute for Neuropathology, RWTH Aachen University, Pauwelsstraße 30, 52074 Aachen, Germany ; Department of Ophthalmology, RWTH Aachen University, Pauwelsstraße 30, 52074 Aachen, Germany.
authors.forename: Florent
authors.initials: F
authors.lastname: Haiss
authors.affiliation: Center for Microscopy and Image Analysis, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland ; Institute of Anatomy, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Dominik
authors.initials: D
authors.lastname: Haenni
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland.
authors.forename: Stefan
authors.initials: S
authors.lastname: Weber
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Marc
authors.initials: M
authors.lastname: Zuend
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Matthew J P
authors.initials: MJ
authors.lastname: Barrett
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Kim David
authors.initials: KD
authors.lastname: Ferrari
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Philipp
authors.initials: P
authors.lastname: Maechler
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Aiman S
authors.initials: AS
authors.lastname: Saab
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Jillian L
authors.initials: JL
authors.lastname: Stobart
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Matthias T
authors.initials: MT
authors.lastname: Wyss
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Helge
authors.initials: H
authors.lastname: Johannssen
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland.
authors.forename: Harald
authors.initials: H
authors.lastname: Osswald
authors.affiliation: Florey Instittue, University of Melbourne, 30 Royal Parade, Melbourne, Victoria, 3010, Australia ; Neuroscience Research Center, Charité-Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany.
authors.forename: Lucy M
authors.initials: LM
authors.lastname: Palmer
authors.affiliation: CSEM Suisse d'Electronique et de Microtechnique, Technoparkstrasse 1, 8005 Zurich, Switzerland.
authors.forename: Vincent
authors.initials: V
authors.lastname: Revol
authors.affiliation: Institute of Anatomy, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Claus-Dieter
authors.initials: CD
authors.lastname: Schuh
authors.affiliation: CSEM Suisse d'Electronique et de Microtechnique, Technoparkstrasse 1, 8005 Zurich, Switzerland.
authors.forename: Claus
authors.initials: C
authors.lastname: Urban
authors.affiliation: Institute of Anatomy, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Andrew
authors.initials: A
authors.lastname: Hall
authors.affiliation: Neuroscience Research Center, Charité-Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany.
authors.forename: Matthew E
authors.initials: ME
authors.lastname: Larkum
authors.affiliation: CSEM Suisse d'Electronique et de Microtechnique, Technoparkstrasse 1, 8005 Zurich, Switzerland.
authors.forename: Edith
authors.initials: E
authors.lastname: Rutz-Innerhofer
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Hanns Ulrich
authors.initials: HU
authors.lastname: Zeilhofer
authors.affiliation: Center for Microscopy and Image Analysis, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Urs
authors.initials: U
authors.lastname: Ziegler
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
authors.forename: Bruno
authors.initials: B
authors.lastname: Weber
keywords: (000.1430) Biology and medicine; (120.4880) Optomechanics; (180.2520) Fluorescence microscopy; (180.4315) Nonlinear microscopy