All microscopes are located on the 6th floor of the OMRF Research Tower.
Confocal Nikon C1 System: This microscope is equipped with three lasers (Ar laser 458, 488, 514 nm, 25 mW; HeNe laser 543 nm, 1 mW; and HeNe laser 633 nm, 5 mW) and filter sets, which allow the use of excitation/emission spectra in the range of FITC, Cy3 and Cy5 fluorochromes.
Zeiss 710 Multiphoton Microscope System: This instrument consists of a high performance optical microscope, the Axio Examiner motorized system equipped with a physiology stage, a Zeiss LSM710 scanning system with nonlinear optics and multiphoton capabilities, and a Chameleon Ultrafast II Ti:Sapphire laser system. The system has two fluorescence de-scanned channels, DIC transmitted light, and a maximum scan resolution of 6144×6144 pixels at 16-bit depth. The microscope is mounted on a vibration-free optical table. The Chameleon® ULTRA II hands-free ultrafast Ti:Sapphire laser, from Coherent Inc., provides femtosecond pulses tuning more than 400nm, from 680nm to 1080nm, with a peak power of >3.3W. In addition, the instrument is equipped with the following five visible confocal laser lines: Ar laser 458, 488, 514 nm, 25 mW; HeNe laser 543 nm, 1 mW; HeNe laser 633 nm, 5 mW. This instrument is used for single and multiple fluorophores, for image acquisition in the X, Y, and Z directions, and for optical serial sectioning in the z-direction (stacks for volumetric analysis). A META Spectral Detector allows separation of fluorophores with significant spectral overlap (i.e. GFP and FITC). The following objective lenses are included: 10x dry (0.3 NA), 25x water (0.7 NA), 40x multi-immersion (1.2 NA), 63x water (1.2 NA), 63x oil (1.4 NA), and 100x oil (1.4 NA).
Fluorescence Life-time Imaging Microscopy (FLIM) for Zeiss LSM 710: This upgrade to the Zeiss LSM 710 multiphoton microscope consists of a time-correlated single photon counting (TCSPC) system produced by Becker & Hickl GmbH. The instrument includes the following components: (i) Simple-Tau 152 TCSPC FLIM—a highly flexible, modular, dual channel parallel acquisition FLIM system; (ii) two HPM-100-40 GaAsP hybrid detector modules, with high count rate, fast response, no after pulsing and no diffusion tail—characteristics that make it perfect both for FLIM and FCS (fluorescence correlation spectroscopy); and (iii) Data Analysis Software for Fluorescence Lifetime Imaging for Windows 95/98/NT/ME/2K/XP/VISTA. The system allows: (i) time-series FLIM as fast as 2 images/second; (ii) FCS recording, online-correlation and fit; (iii) single, double, and triple exponential decay analysis; (iv) FRET measurement without bleed through; (v) double exponential FRET for separation of interacting and non-interacting protein fractions; and (vi) ion concentration measurements.
Spinning-disk Confocal Microscope for Intravital Imaging: Core members have recently set up a state-of-the-art, rapid-scan confocal imaging system that allows fast intravital imaging in small animals (mice, zebrafish, etc.). The instrument is located within an imaging suite along with a wide-field intravital microscope and an intravital multiphoton microscope. We have custom-configured a micro-lensed Yokogawa spinning-disk confocal scan head (CSU-X1). Spinning-disk confocal microscopy allows acquisition of images at very high frame rates with minimum illumination of samples, which makes this technique particularly well suited to high-speed 3D imaging of living systems with low phototoxicity. The system has a high intrinsic scan rate (1000 scans per second) and is mounted on a Nikon Eclipse 600 FN microscope equipped with a physiology stage and water dipping objectives optimized for intravital imaging in small animals. The excitation light will be provided by three laser modules (two solid-state and a diode laser) that give the following wavelength lines: 488, 561 and 640 nm. The system includes an acousto-optic tunable filter (AOTF) and a laser combiner unit that can work with up to 5 solid-state laser modules, thus allowing further expansion of the laser lines as necessary. The AOTF permits the selection of laser lines and intensity modulation and is very important for applications involving photobleaching or photoactivation. The confocal scan head is connected to a 10-position emission filter wheel with 50 ms switching time. Distinct emission spectra are selected using band-pass filters (Brightline single band filters for 525, 607 and 685 nm from Semrock Inc.). The resulting emission light passes through a camera adaptor that connects the filter wheel to the Hamamatsu EMCCD camera. For 3D and 4D (3D plus time) imaging, a Piezo Z objective with 100-μm travel and 60 optical sections per second is included.
Electron Microscope: A Hitachi H-7600 Transmission Electron Microscope equipped with a Kodak 2k x 2k digital monochrome camera and AMT EM image acquisition software is available for Core staff usage in the OMRF Imaging Facility on the ground floor of the OMRF Research Tower on an investigator chargeback basis.