Design and performance of an ultra-flexible two-photon microscope for in vivo research

Johannes M Mayrhofer; Florent Haiss; Dominik Haenni; Stefan Weber; Marc Zuend; Matthew J P Barrett; Kim David Ferrari; Philipp Maechler; Aiman S Saab; Jillian L Stobart; Matthias T Wyss; Helge Johannssen; Harald Osswald; Lucy M Palmer; Vincent Revol; Claus-Dieter Schuh; Claus Urban; Andrew Hall; Matthew E Larkum; Edith Rutz-Innerhofer; Hanns Ulrich Zeilhofer; Urs Ziegler; Bruno Weber

doi:10.1364/BOE.6.004228

Design and performance of an ultra-flexible two-photon microscope for in vivo research

Biomed Opt Express. 2015 Oct 2;6(11):4228-37. doi: 10.1364/BOE.6.004228. eCollection 2015 Nov 1.

Authors

Johannes M Mayrhofer¹, Florent Haiss², Dominik Haenni³, Stefan Weber⁴, Marc Zuend¹, Matthew J P Barrett¹, Kim David Ferrari¹, Philipp Maechler¹, Aiman S Saab¹, Jillian L Stobart¹, Matthias T Wyss¹, Helge Johannssen¹, Harald Osswald⁴, Lucy M Palmer⁵, Vincent Revol⁶, Claus-Dieter Schuh⁷, Claus Urban⁶, Andrew Hall⁷, Matthew E Larkum⁸, Edith Rutz-Innerhofer⁶, Hanns Ulrich Zeilhofer¹, Urs Ziegler⁹, Bruno Weber¹

Affiliations

¹ Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland ; Neuroscience Center Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
² 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.
³ 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.
⁴ Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, 8057 Zürich, Switzerland.
⁵ Florey Instittue, University of Melbourne, 30 Royal Parade, Melbourne, Victoria, 3010, Australia ; Neuroscience Research Center, Charité-Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany.
⁶ CSEM Suisse d'Electronique et de Microtechnique, Technoparkstrasse 1, 8005 Zurich, Switzerland.
⁷ Institute of Anatomy, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
⁸ Neuroscience Research Center, Charité-Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany.
⁹ Center for Microscopy and Image Analysis, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.

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.

Keywords: (000.1430) Biology and medicine; (120.4880) Optomechanics; (180.2520) Fluorescence microscopy; (180.4315) Nonlinear microscopy.