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, immunolabelling procedures and image analysis.
 
The facility supports three imaging systems, the Olympus FluoView FV300 Laser Scanning Microscope, the Zeiss Axiovert 200M Apotome microscope and the Olympus IX70 fluorescence microscope.
 
The FluoView 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, 543 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 facility is in the process of upgrading this microscope to include a CY7 filter and Xenon illumination to allow imaging of IR dyes such as IR800 from LI-COR. The addition of a Fluorescence Resonance Energy Transfer (FRET) module is in progress. This imaging technique 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 Light Microscopy Core Facility hosts imaging-related seminars at the Feinstein Institute and participates in teaching at the Elmezzi Graduate School and the Feinstein Institute community outreach program.

Name:   Amanda Chan Position:   Manager Email:   achan@nshs.edu