Center for Cardiometabolic Science



Envirome Institute

University of Louisville



Cores


The Center houses a suite of core services to support the needs of the investigators in the Center, as well as others on campus.


Services Offered

The Center hosts a broad array of access to instrumentation and procedures. For information about the following areas, please contact those indicated:

SURGERY/CARDIAC FUNCTION

(Contact: Yibing Nong, MD, PhD)
The Center performs animal surgeries (myocardial ischemia with/without reperfusion, transverse aortic constriction, apical resection, intracardiac injection, splenic transplant, radiotelemeter implantation, et al) and measures cardiac function (echo, PV loop and electrophysiology study) for the investigators in the Center, as well as others on campus. This suite of instruments and services also provides robust educational opportunities for trainees and faculty.

VisualSonics Vevo 3100 High Resolution in vivo Imaging System  The 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. 
 Surgery Stations  The Core has five surgery stations. Each station is equipped with a dissecting microscope (Olympus/Nikon); a light source and fiber optics; a mouse ventilator (Hugo Sachs) connected to supplemental oxygen; an automatic, temperature-regulated heat lamp; and a continuously heated water blanket (Braintree Science) to ensure core body temperature is maintained at 36.5–37.5°C. The ECG monitoring system (AdInstruments) or a portable heating and monitoring surgical board (Harvard) can record vital signs of the animal during surgeries.

Telemetry  In addition to traditional telemetry for ECG, blood pressure, or body temperature, the Center also houses a complete 8 channel telemetry system (Data Sciences International, DSI) used to simultaneously measure electroencephalogram (EEG) and electromusculogram (EMG), and perform video monitoring for assessment of sleep/wake cycles and sleep scoring in mice. The system is equipped with 8 HD-X02 implants (two biopotentials, temperature, activity), 8 RPC-1 receivers, 4 video cameras (capable of monitoring 2 cages at a time), Ponemah v6.51, and NeuroScore automated scoring analysis software with Rodent Sleep module. In addition to automated sleep scoring, the system is also capable of seizure detection in rodent models. 
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. It automatically calibrates the volume signal with admittance technology to eliminate the need for hypertonic saline injections and cuvette calibrations. The system is equipped with two catheters for smaller (3.5 mm) or larger (4.5 mm) mice to acquire the most accurate hemodynamic measurements. The LabScribe data acquisition software (LabScribe v3) is used to record and analyze hemodynamic data online/offline.
Electrophysiology Study System  The Center has a Programmed Electrical Stimulation (PES) system for evaluating refractoriness, nodal recovery time, and inducibility of arrhythmia in the ventricular and atrial myocardium in vivo or ex vivo.  The system consists of an STG-3008 combined stimulator and amplifier (MultiChannel Systems) and Powerlab matrix with ML136 Animal Bio Amp operated with LabChart Pro 8.0 software (ADInstruments). The STG-3008 is connected to a 1.1-French 8-electrode octopolar catheter either alone (EPR-800, Millar) or bearing a lumen for drug or test agent delivery (EPR-801, Millar). The octopolar catheter facilitates programmed region-specific stimulation of cardiac tissue and concurrent monitoring of intracardiac electrophysiology across the different chambers of the beating heart.

MICROSCOPY/PATHOLOGY

(contact: Matthew A. Nystoriak, PhD)
Nikon A1R-HD Confocal/Multiphoton Microscope  This is a turn-key, confocal, multiphoton system that includes a high-speed resonant scanner with a large field-of-view and high-resolution scanning options for live cell or intravital imaging. It also has a traditional Galvano scanner for high spatial resolution needs. The system is mounted on a FN1 upright microscope body with a motorized XYZ platform and a high-speed piezo z drive system for fast z-stack acquisitions and is equipped with 4 visible laser lines (405, 488, 561, 640 nm) and GaAsP detector system, and a Coherent Discovery dual beam IR laser. The system currently has 7 objectives. The system has a high-end image acquisition workstation with NIS-Elements software as well as an additional workstation dedicated for offline image analysis.

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 Center 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.

Bruker JPK NanoWizard 4 XP BioScience Atomic Force Microscope  This system reflects the state-of-the-art capabilities for interrogating tissue mechanics. The system has scanning rates up to 150 lines/sec and up to 100 µm scan range. This system is suitable for living cells/tissue and fresh tissue. The system can also perform automatic overlay with transmitted light images and fluorescence microscopy images. In addition, this AFM scan head can quickly be moved to other diascopic, confocal, or epifluorescent systems. Although this system is configured with a dedicated Zeiss microscope, the Bruker AFM scanhead could be redeployed to the Nikon A1 confocal or Nikon A1R multi-photon confocal system, as needed.
Full Pathology Suite  The Center has a complete Sakura pathology suite (automated processor, embedder, microtome, and cryostat). This, like all self-service elements in the Center, can be scheduled via the electronic scheduling system (see bottom of page).

EXERCISE/METABOLIC CAGES

 (contact: Jason L. Hellmann, PhD)
Exercise Systems  The Center has three Columbus Instruments six-lane rodent treadmills with software-controlled capabilities. The Center also maintains 50 in-cage voluntary exercise mouse running wheels equipped with VitalView activity software for 24 h automated recording. In addition to forced and voluntary exercise systems, the Core maintains two 15-gallon tanks for swimming exercise in mice.

Metabolic/Behavioral Phenotyping System The Center has an 8-cage behavioral and metabolic phenotyping Promethion Core system (Sable Systems International) outfitted with voluntary wheels and programmable access to food and water. The system is equipped with a stable isotope gas analyzer for analysis of nutrient utilization and the system operates within an environmentally controlled cabinet with programmed passcode protected accessibility. Using Promethion Live software, real-time data for each parameter of every cage can be viewed, allowing users to make key decisions at any time, from any place. We also have a complete TSE PhenoMaster/LabMaster Metabolic Chamber system to monitor individual mouse metabolic functions including indirect calorimetry (O2 consumption, CO2production, RER), food intake, water consumption, and total activity (ambulatory and fine motor activity in X-axis).

EXPOSURE STUDIES AND TOXICOLOGY

(contact: Daniel J. Conklin, PhD)
Portable GE Medical Systems Lunar PIXImus Dual-Energy X-Ray Absorptiometer (DEXA) Scan  The Lunar PIXImus Densitometer (GE Medical Systems) provides bone mineral and body composition (lean and fat body mass) composition results from total body imaging in less than 5 min. 
AVOXimeter 4000 Whole Blood CO-oximeter  Performs individual measurements of total hemoglobin (tHb), oxyhemoglobin saturation (%O2Hb), carboxyhemoglobin (%COHb), and methemoblobin (%MetHb) on freshly-drawn or heparin- or EDTA-anticoagulated whole blood samples as small as 50µL. Oxygen content ([O2Ct]), percent saturation (SO2) and oxygen carrying capacity (O2Cap) of the blood sample are automatically calculated from the %HbO2 and THb measurements.
Kent Scientific CODA 6 Non-invasive Blood Pressure Monitoring System  A computerized system that automatically performs rapid, simultaneous, multiple measurements of six physiological parameters: systolic blood pressure, diastolic blood pressure, mean pressure, heart rate, tail blood flow, and tail blood volume, on up to six test subjects.

Inhalation Facility  This barrier style facility houses an AAALAC-certified animal room (500 ft2) and a procedure room consisting of 240 ft2 of laboratory space, a biosafety cabinet, fume hood, microbalance, refrigerator, and 4 linear feet of dedicated bench space. Exposure laboratories comprise two rooms (380 ft2 and 200 ft2) equipped with aldehyde and cigarette smoke exposure systems. A separate vapor exposure system delivers volatiles from either permeation tubes (Kin-Tek; LaMarque, TX) or concentrated gas tanks (AirLiquide, Plumstead, PA) for volatile compound exposures with real-time monitoring using RAE Systems detectors (PID; ppbRAE Plus; MultiRAE). For smoke exposures, there is a complete 4-chamber inExpose system (SCIREQ; Montreal, CAN) robotic cigarette smoke system capable of performing multi-dose level exposures using programmable mixing of mainstream and sidestream smoke. The Facility also houses a Versatile Aerosol Concentration Enrichment System (VACES) with capacity to simultaneously expose 64 mice to both ambient concentrated particulate matter (CAPs) and to HEPA-filtered air.

ADDITIONAL SERVICES

Flow Cytometry  This lab is fully equipped with a state-of-the-art cytometer (described below) and analysis software. This BD Fortessa flow cytometer is equipped with 4 lasers: a 488nm (2-color detection), a 640nm (3-color detection), a 405nm (6-color detection), and a 561nm (4-color detection). It also has high throughput capability (contact: Jason L. Hellmann, PhD).
Analysis of Live Cellular Metabolism  The Core also houses an extracellular flux analyzer (Agilent XFe96) 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 insights into metabolic function (contact: Bradford G. Hill, PhD).




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