The Vascular, Intravital, MulTiphoton, and Advanced AnaLytics (VITAL) Facility is a multi-user equipment and analysis core facility designed to provide microvascular and vascular scientists with the advanced technologies necessary to study all aspects of the circulation, including its structure and delivery of nutritive blood flow, in live animals or in ex vivo preparations and immunohistochemistry. While built around vascular imaging, the equipment is broadly applicable to a range of intravital and tissue imaging approaches.
Location & Contact
Location: Room 3277, Robarts Research Institue
Facility Director: Dr. JG Pickering, gpickering@robarts.ca
Facility Manager: Stephanie Milkovich, smilkovi@uwo.ca
Available Instruments
Nikon A1R MP+ Multiphoton Upright Microscope
The Nikon A1R MP upright multiphoton microscope can be used to capture fluorescent images deep into tissues. Cells and live organs can be imaged at a greater depth than is currently possible with typical laser-scanning confocals. Non-intravital applications for multiphoton microscopy include tracking fluorescent protein-expressing immune cells and visualizing the dynamics of drug- and metal-containing nanoparticles.
Housed within a dark enclosure, a height-adjustable motorized XY stage can accommodate multiple model organisms and live animals, in addition to cultured cells. The tunable long wavelength IR laser allows for simultaneous imaging of a variety of fluorescent probes.
Nikon Ti2-E Confocal Inverted Microscope
The Nikon Ti2-E Inverted Microscope paired with the A1R HD laser-scanning confocal offers an 18 mm field of view; which means more data per image, less stitched images and fewer artifacts. With its real-time hardware focus correction and intuitive/user-friendly operation, the A1R HD is perfect for imaging both fixed tissue, live cell, and intravital preparations.
Interference filters and a 80-20 split mirror enable measurement of RBC oxygen saturation and flow parameters in microvasculature using absorption spectroscopy. Alpha Mass Flow Controllers allow changes in oxygen levels in a custom-chamber under the intravital preparation to challenge microvasculature. Responses to such changes are measured via changes in flow or with perictyes in the tissue.
Surgical Equipment
A range of small animal surgical equipment is available to support intravital microscopy. This includes anesthesia and monitoring systems, dissection microscopes, laser doppler imaging, and blood spectrophotometry.
Analysis
The VITAL Core Facility offers our users four workstations for image analysis. Acquired data images are accessed via secure server for analysis using Nikon NRS Software Analysis and an additional Imaris workstation for generic image analysis.