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 (MI with/without reperfusion, TAC, apical resection, intracardiac injection, splenic transplant, and et al) and measures cardiac functions (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.

picture of Vevo 3100 station
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. 

picture of surgery station

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. 
picture of PV loop system

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.
picture of PES system

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)

picture of confocal microscope
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.

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).

MASS SPECTROMETRY

(contact: Paweł K. Lorkiewicz, PhD)

picture of Synapt

picture of Sciex 5500


Currently, our Center has excellent targeted analytical workflows based on the state-of-the art UPLC/UHLPC-MS/MS systems with triple-quadrupole analyzers. In August 2020 the Center added a high resolution, state-of-the art Waters Synapt XS LC-MS system (obtained through the NIH shared instrumentation grant mechanism – S10 OD026840). We facilitate characterization and quantitation of small molecular weight analytes in biofluids, tissues and cells. Services include:
  • UPLC-MS/MS measurements
    • 28 VOC metabolites and 5 tobacco alkaloids in urine
    • Low abundance VOC metabolites in urine
    • Biogenic monoamines and their metabolites in urine
    • Histidine/histidine dipeptides in tissues/urine
    • Measurement of cortisol/cortisone in urine
    • Muconic acid in urine
    • LC-MS/MS analyses of 41 lipid mediators
  • Stable isotope resolved metabolomics analysis - data acquisition (Synapt XS) and analysis
  • Discovery and identification LC-MS
    • Untargeted and semi-targeted metabolomics analyses
    • Screening and identification of xenobiotic metabolites (mercapturates, GSH-conjugates, 1st and 2nd phase metabolites)
    • Analyte confirmation (accurate mass, fragmentation)
  • Method development
    • Synapt XS untargeted and targeted method development
    • LC-MS/MS quantitative method development
    • GC-MS method development (instrument currently unavailable)
  • Training

EXERCISE/METABOLIC CAGES

 (contact: Jason L. Hellmann, PhD)
The murine exercise and metabolic monitoring suite contains two six-lane treadmills, 24 computer-monitored voluntary wheels, and a Sable 8-cage metabolic cage monitoring system (with exercise wheels).

EXPOSURE STUDIES AND TOXICOLOGY

(contact: Daniel J. Conklin, PhD)

picture of DEXA scanner

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. 
picture of metabolic chamber system
TSE PhenoMaster/LabMaster Metabolic Chamber System - A highly-flexible modular system for state-of-the-art automated metabolic and behavioral monitoring of small rodents. It is located in a dedicated, environmentally-controlled room (12h:12h light:dark cycle; red light equipped for afterhours treatment; 150 ft2).
picture of AVOXimeter
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.
picture of blood pressure monitoring system
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.
picture of telemetry system
Telemetry System - Uses implantable telemetry technology to collect data in real time to monitor body temperature, activity, blood pressure, respiratory function, ECG, and blood glucose.

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 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 insights into metabolic function (contact: Bradford G. Hill, PhD).