Design and installation of a multimode microscopy system

Abstract

We describe design and installation of a multi-mode microscopy core facility in an environment of varied research activity in life-sciences. The experimentators can select any combination of a) microscopes (upright, upright fixed-stage, inverted), b) microscopy modes (widefield, DIC, IRDIC, widefield epifluorescence, transmission LSM, reflection and fluorescence CLSM, MPLSM), c) imaging techniques (direct observation, video observation, photography, quantitative camera-recording, flying spot scanning), d) auxiliary systems (equipment for live specimen imaging, electrophysiology, time-coordinated laserscanning and electrophysiology, patch-clamp). The equipment is installed on one large vibration-isolating optical table (3m × 1.5m × 0.3m). Electronics, auxiliary equipment, and a fiber-coupled, remotely controlled Ar + -Kr + laser are mounted in a rack system fixed to the ceiling. The design of the shelves allows the head of the CSLM to be moved to any of the microscopes without increasing critical cable lengths. At the same time easy access to all the units is preserved. The beam of a Titanium-Sapphire laser, controlled by means of an EOM and a prism GVD, is coupled directly to the microscopes. Three mirrors mounted on a single precision translation table are integrated into the beam steering system so that the beam can easily be redirected to any of the microscopes. All the available instruments can be operated by the educated and trained user. The system is popular among researchers in neuroanatomy, embryology, cell biology, molecular biology -including the study of protein interactions, e.g. by means of FRET, and electrophysiology. Its colocalization with an EM facility promises to provide considerable synergy effects.

Structural changes at the cellular and subcellular level in the cerebral cortex of mice visualized by means of trans-cranial multi photon in-vivo microscopy

“…The new setup is integrated into, and has replaced one setup on a large frame facility described earlier 6 . The core of the setup is an upright microscope with a focusing objective revolver and a scan unit, into which the beam of a Ti:Sap laser is guided directly, i.e.…”

Analysis of optical properties of the mouse cranium—Implications for in vivo multi photon laser scanning microscopy

“…The scanning unit is equipped with a transmitted light photomultiplier tube (PMT, model R6357, Hamamatsu Photonics K.K., Hamamatsu City, Japan). (IV) The galvanometric z-stage of the microscope is in itself fixed to a home made microscope stage (Helm et al, 2001), which allows to position the preparation in x, y and z controllably at a precision of better than 1 m. (V) The preparation can be moved relative to the objective and condenser at which objective and condenser do not move relative to each other. This, of course, implies that the condenser can be moved for focussing purposes and adjustment of proper Köhler illumination while the stage and preparation remain totally un-touched.…”

“…The readings of the three mechanical actuators were recorded. Then, using the mechanical z-drive of a precision microscope stage (Helm et al, 2001), the stage was lifted by 125 m (see "A" in Fig. 1 V).…”

Optical effects of the cranium in trans-cranial in vivo two photon laser scanning microscopy in mice

“…The beam of the Ti:Sap laser was controlled by means of a model "LM 0202P 5W IR / LIV8" Electro Optic Modulator system (Gsaenger / Linos, Planegg, FRG). The setup has been described in detail in earlier publications 5,10 . The respective software modules used for the data acquisition were "LCS, Vs. 2.5 Build1347" (Leica Microscystems) and "Fluoview, Vs. 5.0 Build 92" (Olympus).…”