Imaging and Physiology
The Imaging and Physiology Core promotes scientific excellence in investigations of cardiometabolic disease. It provides investigators with access to state-of-the-art instrumentation and expert instruction related to confocal imaging, assessment of cardiac function, and performance of surgery in mice.
The Core will promote education on the theory, as well as sound technical comprehension, related to the various instruments and approaches available. This Core enables the integration of biochemical insights at the level of intact cells, organs, and intact organisms.
Aims of the Core
- Support current COBRE and Center projects. The Director is dedicated to monitoring the specific needs of Center members as they may relate to confocal imaging, in vivo assessment of cardiac function, and surgical models.
- Educate and train investigators. Individuals with ongoing needs for specific pieces of instrumentation will not simply be consumers of the Core’s resources. Instead, they will be investigators versed in thoughtful application of a particular imaging or physiological modality.
- Develop new protocols and techniques. The Core has developed new techniques. All of these advances were brought to the Core to promote the investigations of the Center members, particularly the Center's early career investigators.
Facilities & Instrumentation
VisualSonics Vevo 3100 High Resolution in vivo Imaging System
The Diabetes and Obesity Center houses a VisualSonics Vevo 3100 High Resolution in vivo Imaging System (photo, right). This system is a non-invasive ultrasound-based method for viewing extremely small physiological structures in mice.
This real-time, in vivo imaging system has a spatial resolution down to 30 microns - the highest resolution available - and provides advanced imaging modules such as 4D and strain. .
Left Ventricular Catheterization System
The Core is also equipped with an ADVantage Pressure Volume System (ADV 500) single-segment pressure-volume(P-V) conductance and admittance system (Transonic). This system can simultaneously and continuously measure left ventricular pressure and volume in intact small animals.
Confocal Imaging Laboratory
Nikon A1 laser scanning confocal microscope
The confocal imaging laboratory houses a state-of-the-art Nikon A1 laser scanning confocal microscope (photo, right). This system is mounted on a customized, inverted, fully electronic, TE-2000E2 microscope body.
This system offers unparalleled spatial resolution in the x, y, and z planes.
There are four excitation lines of 405, 488, 561, and 640nm, which are driven by four (4), independent solid-state lasers (superior stability and longevity) mounted on an acousto-optic tuning filter (AOTF)-integrated laser launch.
Automated Fluorescence Microscope
The Core recently acquired a state-of-the-art, all-in-one, fluorescence microscope BZ-X810 (Keyence). This microscope offers the imaging capabilities of a traditional fluorescence microscope with additional capacities for image stitching and z-stacking, optical sectioning, and automated time-lapse imaging.
Analysis of Live Cellular Metabolism
The Core also houses two extracellular flux analyzers- an Agilent XF96e and a Seahorse XF24 - to measure metabolism in live cells, small organisms, and tissue explants. In addition, these instruments can be used to measure electron transport chain activity in isolated mitochondria or permeabilized cells. These systems measure oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), which are indicators of mitochondrial respiration and glycolysis, respectively. These measurements provide insight into how metabolic function goes awry in disease states or changes with physiological stressors or genetic manipulations.
In collaboration with Martha Bickford, Ph.D. (KBRIN Director), the Core offers assistance in sample TEM sample preparation and an orientation to TEM imaging. For more details please see the KBRIN website.
Additional Areas of Expertise
- Helen E. Collins, Ph.D. (electron microscopy; calcium/contractility imaging; general histology)
- Bradford G. Hill, Ph.D. (mitochondrial function; perfused heart; SIRM data analyses)
- Steven P. Jones, Ph.D. (cardiac function; ultrasound imaging; microscopy)
- Tamer Mohamed, Ph.D. (cardiac MRI; high-throughput imaging and analysis; microscopy)
- Joseph B. Moore, IV, Ph.D. (induced pluripotent stem cells; cell-lineage differentiation)
- Matthew N. Nystoriak, Ph.D. (vascular function/blood flow regulation; microscopy)
- Marcin Wysoczynski, Ph.D. (immune cell physiology and cytometry; bone marrow chimera models)