SCHOOL OF MEDICINE

Department of Medicine

Indiana Center for Biological Microscopy

Current Facility

Olympus FV1000 MPE

This microscope is configured for both multiphoton and standard confocal imaging and can perform FRET and FRAP experiments.  It is an inverted system providing confocal laser excitation at 405, 458, 488, 514, 559 and 635 nm and multiphoton fluorescence excitation provided by a fully automated Spectra-Physics MaiTai DeepSee laser. A variety of standard and long working distance high resolution water and oil immersion objectives are available for use on this microscope.   The system is ideally suited to intravital microscopy and can be equipped with an objective heater, heating pads and a thermal blanket.

Olympus FV1000 confocal/home-built multiphoton

This is a confocal/home-built 2-photon microscope mounted on an inverted microscope stand. It offers confocal laser excitation at 488, 514, 568 and 633 nm and multiphoton laser excitation at 690-1040nm provided by a Spectra-Physics MaiTai DeepSee tunable laser. The system is equipped with three internal detectors including two spectral detectors and a transmitted light detector. In non-descanned mode it offers three-channels acquisition, supporting increased signal-to-noise ratio imaging provided by two GaAsP (Gallium Arsenide Phosphide) detectors. In addition, the system is equipped with a motorized stage supporting mosaic and registered fields acquisition. It is ideally suited to a regular fixed samples visualization as well as an intravital microscopy. It can be equipped with an objective heater, heating pads and a thermal blanket and offers a variety of standard and long working distance high resolution water and oil immersion objectives.

Spinning disk confocal and live-cell microscope

This microscope is configured for high-speed live cell imaging of fluorescent proteins, FRET and immunostained samples in three dimensions.   Built around a Nikon TiE inverted microscope stand, it is a combined confocal and widefield microscope system.  Confocal microscopy is based upon a Yokogawa spinning disk, with laser illumination provided at 442, 488, 514 and 561 nm via an Agilent launch with images collected by an Andor EM-CCD.  Widefield microscopy can be conducted using a Xenon lamp source, optics suitable for excitation from 340 nm to 700 nm, with images collected by a Hamamatsu interline CCD.  It is equipped with a Perfect Focus system, a motorized stage and a stage-top incubator for maintaining a heated and CO2 dependent environment to ensure stable long term imaging of cultured cells.  Heated high-resolution water and oil immersion objectives are available on this microscope including fluorite lenses for imaging below 400 nm.

Leica SP8 MP

This microscope is configured for both multiphoton and standard confocal imaging.  It is an upright motorized Leica SP8 MP with a fixed large-platform motorized stage equipped for confocal excitation at 405, 448, 488, 552 and 633 nm with mulltiphoton fluorescence excitation provided by a fully automated Spectra-Physics MaiTai DeepSee laser.  The SP8 MP includes both standard galvanometric scanning for imaging at up to 1800 Hz and resonant scanning for imaging up to 8000 Hz, supporting full frame imaging at up to 30 fps.  The system is ideally suited to intravital microscopy and can be equipped with  an objective heater, heating pads and a thermal blanket.

OpenSPIM

This microscope is a light-sheet microscope based on the openSPIM design that can image large samples with isotropic high-resolution through multiview imaging.  The base L-SPIM design was extended to include additional excitation wavelengths and support for live imaging. It uses an ORCA Flash 4.0 v2 using CameraLink for fast (100 fps) and large area imaging.  To support imaging of fluorophores across the visible spectrum it has been fitted with laser excitation at 442, 488, 514, 568 and 647 nm and automated emission filters.  Live imaging is made possible with automated control of the chamber’s temperature (25-42°C) and faculties for perfusion/media recirculation.  A powerful workstation is provided to facilitate image capture and processing including the acquisition of multi-terabyte datasets and 128 GB of RAM and a pair of nVidia GPUs to facilitate processing of multiview image fusion and deconvolution.