Intravital Imaging Core
Core Director
Dr. Jensen Smith is a Research Assistant Professor at UNMC. Her scientific contributions have focused on the development and use of various imaging methodologies for representation of biological phenomena during normal and disease states. Her particular expertise has been in studying the cellular mechanisms of hearing loss. She oversees the multiphoton microscope and assists investigators in use of intravital microscopy and multiphoton excitation/imaging to collect both high-resolution and whole brain images in live animals, fresh tissue explants, as well as various types of fixed samples.
Core Equipment
Contact Dr. Jensen Smith for more information or for scheduling.
Multiphoton Microscope
Olympus FVMPE-RS microscope, a multichannel, multiphoton excitation system specifically tailored for brain imaging. It delivers high speed (up to millisecond imaging) for rapid in vivo imaging of physiological responses, near infrared excitation for deep tissue imaging. The Olympus FVMP-RS system is additionally equipped with 458 and 588 nm stimulation laser lines for photobleaching, light stimulation and/or photoactivation of various fluorescent or caged constructs. A Spectra-Physics InSight X3 dual-line multiphoton laser offers ultrafast, multichannel excitation and imaging (1045nm fixed, 680-1300nm tunable). The extended excitation range excites both traditional fluorophores (blue to long range red) and can be used for second and third harmonic imaging. The Neurotar Mobile Home Cage allows researchers to integrate real-time imaging with behavioral/locomotion tracking.
Cost: $60/h for assisted microscopy. Users accruing significant experience and training during assisted imaging sessions are eligible for unassisted imaging ($30/h). Unassisted imaging is approved at the discretion of Dr. Jensen-Smith.
Light Sheet Fluorescence Microscope
Tissues are inherently three dimensional in nature, which makes imaging intact tissues a necessity for a more complete study in the relationship between structure and function and the system-level study of cellular mechanisms. The Miltenyi Biotec (previously La Vision) Ultramicroscope II Light Sheet fluorescence microscope (LSFM) extends your existing fluorescent imaging into true 3D, large-scale volumetric imaging the nervous system. Optimized volumetric imaging requires removal of light bending cellular components (mostly lipids) through a process called optical clearing.
Cost: $40/h for assisted microscopy. Users accruing significant experience and training during assisted imaging sessions are eligible for unassisted imaging with technological assistance available ($30/h) and/or unassisted imaging without technological assistance ($20/h). Unassisted imaging options are approved at the discretion of Dr. Jensen-Smith.
X-Clarity™ System
Tissue clearing is important for 3D imaging of tissues at single-cell resolutions. The X-CLARITY™ system simplifies and accelerates each step of the tissue clearing process using CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging / Immunostaining /in situ-hybridization-compatible Tissue hYdrogel). Native cytoarchitecture remains intact and endogenous fluorescence proteins are preserved for robust fluorescence imaging downstream. For LSFM imaging, samples should be optically cleared (X-clarity), labeled with antibodies, then equilibrated in refractive index (RI) matched mounting solutions for imaging.
Cost: $100/run. Each run includes hydrogel embedding and subsequent clearing. Researchers are responsible for subsequent antibody labeling (if applicable) and refractive index matching prior to imaging. Large sample may require extended clearing ($150/ long run).
Imaris-Equipped Workstation
IMARIS is the world’s leading Interactive Microscopy Image Analysis software focused on 3D and 4D imaging. The complete IMARIS for Core Facilities software package with focused image analysis, segmentation, and reconstruction options for cancer and neuroscience researchers, cell and developmental biologists is available for use. IMARIS can be used for any number of imaging studies but is anticipated to be particularly relevant for researchers using the Ultramicroscope II light sheet fluorescence microscopes.
Cost: $5/h unassisted, $40/h assisted.