The University Animal Care facility maintains and manages AAALAC approved animal facilities. Programs of animal husbandry, preventive medicine, pre- and post-surgical care have been developed to assure adequate veterinary care is provided at all times. Complete veterinary, diagnostic, and clinical support
services are available.
The STBL has 11 personal computers, each with 3Ghz processors, a 500GB hard drive, and 2G memory. There are also four instrument computers in the STBL. These computers run software for digital image processing, the biaxial devices, a small angle light scattering device, and the uniaxial tensile testing device.
All computers will be made available to perform the research proposed in this proposal.
Image Processing Computer:
The STBL has a 40 core 2 node image processing PC with 500Gb of RAM and 1TB of hard drive space for use as an image processing and computational simulation resource.
Advanced Intravital (Multiphoton)
The Vande Geest Laboratory oversees, maintains, and utilizes an advanced
intravital multiphoton microscope within the Center for Biotechnology. This
microscope is within the same building as the PIs main and cell culture
laboratory and is capable of:
Micro-Optomechanical Device (MOD)
– Planar biaxial and pressure inflation configurations
We have designed, tested, and published manuscript utilizing this MOD for ocular, mouse aortic, and tissue engineered vascular constructs. The MOD fits under the multiphoton microscope (described above) and thus allows simultaneous collagen and elastin fiber
mapping of tissues during deformation.
Large scale planar biaxial tensile testing device
Sequential Digital Imaging Correlation of Scleral and Optic
Intravital Imaging Vascular Tissue Engineering
Over the past two and half years the PIs laboratory has designed, fabricated, and tested a novel vascular tissue engineering bioreactor that is capable of imposing axial stretch (with measured axial load) and intraluminal flow and pressure to cellularized tissue
engineered vascular constructs. This bioreactor utilizes a Harvard Apparatus PID feedback controller to run three simultaneous pumps in concert with three inline pressure transducers, allowing controlled cyclic intraluminal pressures, with peak pressures up to 300 mmHg and pulse pressures (systolic – diastolic)
ranging from 10 to 60mmHg. The most novel aspect of these bioreactors, however, is how that they were specifically designed to fit underneath the two photon advanced intravital microscope described above (see lens objective cup in the below figure), without loss of sterile conditions. These bioreactors will allow
the PI and his research team to monitor ECM remodeling (collagen/elastin organization) and nondestructive protease activity (via proteolytic beacons, see Haskett et al. 2016) at any point during culture of our vascular constructs. As seen in the Figure below, the PIs laboratory can currently test
three TEVG constructs at the same time with these three bioreactors. The current research proposal has budgeted for three additional bioreactors, which will allow us to run six TEVG cultures simultaneously. These bioreactor are also large enough that additional samples not requiring axial stretch can be
run within the culture chamber with exposure to the same extraluminal culture medium.
Small Angle Light Scattering Device
The STBL is equipped with a Nikon 90i fully automated (nosepiece, condenser, objectives, fluorescence, z motor, xy stage) upright microscope. The microscope has 4 plan apochromat air objectives (2X NA 0.10 WD 8.5mm, 4X NA 0.20 WD 20mm, 10X NA 0.45 WD 4mm, 20X NA
0.75 WD 1mm) with slots for two more objective lenses. Images are captured with either a 12 bit 20 MHz Digital Monochrome camera, or a 32 MP color CCD camera with the capabilities to perform binned or unbinned acquisition in still picture or video. The microscope has the
capabilities to perform epifluorescent imaging of dyes in the ranges of FITC, TRITC, Cy3, and Cy5. Standard Hg illumination is also possible.
Culture and Gene/Protein Work: