Diseases such as cancer and neurodegeneration on the cellular level can be reflected in aberrant migratory behavior or perturbations of cellular organization and structure. The Light Microscopy Core Facility provides state of the art optical imaging technologies that allow researchers to obtain information on the impact of pathologies on these cellular features. The facility provides support for investigators with design and troubleshooting of protocols, in addition to expert technical assistance with sample preparation, immunolabeling procedures and image analysis. Facility equipment is available to all trained users 24/7 on an unassisted basis. New users will be trained on microscopes between 9:00 AM and 5:00 PM.
The facility supports five imaging systems, the Olympus FluoView FV1000 Multi-Photon Microscope, Olympus FluoView FV300 Confocal Laser Scanning Microscope, Zeiss Axiovert 200M Apotome microscope, Olympus IX70 fluorescence microscope and a Leica LMD7000 Laser Microdissection System.
The FluoView FV1000 Multi-photon is a Laser Scanning Microscope that allows fluorescence imaging deep within specimens. Utilizing a pulsed IR laser, the multi-photon microscope is able to image hundreds of microns into a specimen with minimum phototoxicity and photobleaching. This system has a Spectra-Physics Mai-Tai IR laser tunable between 610 nm to 1020 nm, 4 channel descanning detectors and a motorized stage with capability for live cell, tissue and animal imaging as well as a set up for electrophysiology studies.
The FluoView FV300 microscope allows basic imaging to advanced techniques in a confocal mode. Software-controlled shutters and continuously variable neutral density filters positioned independently before each individual laser, allows the selection of any combination of laser line excitation and light intensity while optimizing simultaneous or sequential multi-channel imaging. The instrument has multiple lasers producing the following excitation wavelengths 488 nm, 543 nm and 633 nm, respectively exciting FITC, Rhodamine and CY5.
The Zeiss Axiovert 200M system enables researchers to capture and analyze fluorescence (from dyes including DAPI, FITC and Rhodamine) and phase contrast images with a black and white Zeiss Axiocam MRM cooled CCD camera. Color images can be captured with a Zeiss Axiocam MRC cooled CCD camera. The Zeiss Axiovert 200M is equipped with an apotome that allows optical sectioning. The addition of a Fluorescence Resonance Energy Transfer (FRET) module allows investigators to study protein-protein interactions, permitting high resolution co-localization studies, and biosensors that report on the activation state of proteins.
The IX70 system enables researchers to capture and analyze fluorescence and phase contrast images with a highly sensitive camera (Orca II cooled CCD camera, Hamamatsu). The instrument is a widefield florescent microscope equipped with DAPI, FITC, Rhodamine and CY5 excitation filters. The IX70 system can be used for still images of fixed samples on coverslips or multi-well plates and time-lapse imaging of live cells in 35 mm dishes. Live cell imaging allows investigator to track cell migration, cell apoptosis and additional features. Image capture and analysis is performed by the Esee and Isee software packages (Inovision, Raleigh, NC).
The Leica LMD7000 Laser Microdissection System enables investigators to isolate very small clusters of cells from a thin tissue section. This method is most often used to obtain DNA/RNA/protein from single cells or a small group of cells within a tissue. The Leica LMD7000 Laser Microdissection System is equipped with a CCD camera with a motorized stage and AVC standard software that allows for automated cell recognition and cutting. This system can be used on fixed tissue or live cells with brightfield or epi-flourescence imaging modules.
The Light Microscopy Core Facility hosts imaging-related seminars at the Feinstein Institute and participates in teaching at the Elmezzi Graduate School of Molecular Medicine and Donald and Barbara Zucker School of Medicine at Hofstra/Northwell.
Amanda Chan, PhD