Center for Hypertension and Precision Medicine

The physiological core is well equipped with various instruments to provide technically advanced physiological evaluation in rodents.
Cardiac and vascular functional assessments: Two highly sophisticated 4-channel multi myograph systems available to measure vascular reactivity (Multi wire myograph system - 620M, Danish myotechnology, Denmark); A Millar pressure-volume catheter and its recording system, to measure PV-loops and dP/dT for diastology and cardiac function; Acuson Sequoia™ C512 Ultrasound system for echocardiography from SIEMENS (This item is available through the University core laboratory facility); Mouse Monitor S heated platform with pulse oximeter module and ultrasound cable to sync to Asucon sequoia C512 for ECG and pulse oximetry from Indus Instruments.

Blood Pressure measurements: Two IITC equipment for semi-automated tail cuff blood pressure measurement in rats and mice, two CODA tail-cuff blood pressure system from Kent scientific and apparatus for direct blood pressure measurements of 24 animals concurrently by telemetry (Data Sciences International). Apparatus for direct blood pressure measurements of 16 animals concurrently by wireless stellar telemetry (TSE).

Surgical equipment: Schott ACE I goose neck fiber-optic light source for visualization during surgical and other procedures; Kent Scientific SurgiSuite Heated surgical platform with LED light and magnetic retractor sets for mouse and rat; Kent Scientific mouse/rat intubation kit and stand; Harvard Apparatus MiniVent type 845 rat/mouse ventilator; Olympus SZX7 Dissecting microscope with Olympus DF PLAPO lens and a boom stand; Dage MTI color HD camera with connections for recording pictures and video.

Renal function measurements: Two metabolic cage units to collect urine samples from 72 animals.  NanoDrop 3300 Fluorimeter from Thermo Scientific for FITC-inulin based measurement of whole animal and single kidney glomerular filtration rate (GFR).

Aerobic running capacity tests: Two rodent treadmills to measure the aerobic running performance of the rats from Columbus Instruments (Model Exer-4; Columbus, OH).

The genomic core contains equipment required for genotyping, functional assays, mutation detection and positional cloning.StepOnePlus™ 96-well Real-Time PCR System from AB-bio system; digital camera for recording agarose electrophoretograms from Gel logic- 100 imaging system, Kodak. Protean II Xi 2-D Electrophoresis from Bio-Rad; Omega Lum G chemiluminescence/UV-VIS gel and membrane documentation system from Aplegen; Blotcycler automated membrane processor from Precision Biosystems; Applied Biosystems (ABI) 7500Fast qPCR machine; Qiacube from Qiagen-for extraction of RNA, DNA and Protein; TissueLyser LT with 12 tube adaptor from Qiagen; Agilent 2100 Bioanalyzer. Sequencing services are centrally negotiated through MWG.

The laboratory of vascular pathology consists of a dedicated area (~100 sq. ft.) of laboratory space fully equipped for animal surgery, dissection, necropsy and cryosectioning. The pathology laboratory is adjacent to a cold room and refrigerators and freezers, and is equipped with a Zeiss Axiovert 40CL inverted microscope (bright field and fluorescence, with X-Cite 120Q Illuminator system) with Micropublisher 3.3 Megapixel Cooled CCD Color Digital Camera, coupled to NIS Elements D software for acquisition and analysis, including camera and Nikon Motorized Microscope Drivers and computer system; a HM550 Cryostat (Thermofisher) and dissection microscope with stroboscopic lights. A separate area within the pathology laboratory is dedicated to tissue and slide processing for immunohistochemistry and immunofluorescence. 

The lipidomics core is operated and maintained by Dr. Levison, who is an expert in the use and operation of high end analytical instrumentation including various High Performance Liquid Chromatographs and Mass Spectrometers. The facility includes Single Quadrupole and Ion Trap GC/MS, High Resolution Q-TOF and Orbitraps, Triple Quadrupoles and Quadrupole Ion Traps. These LC/MS systems provide the necessary chromatographic resolution to optimize sensitivity and specificity for the identification of disease pertinent bio-markers and drug metabolites. Dr. Levison expertise in liquid chromatography specific to high throughput analyses includes online solid phase extraction, and multiplexed multichannel parallel HPLC.

The biochemical core contains all the standard equipment necessary for all biochemical analysis from tissue/cell culture samples. Polytron- PT3100D tissue homogenizer from Kinematica AG, temperature controlled Sorvall RC-5C Plus Superspeed Centrifuge, NanoDrop 2000 UV-Vis Spectrophotometer from Thermo scientific, SpectraMax spectrophotometric micro plate reader from Molecular Devices and GloMax-96 Microplate Luminometer from Promega. Two laminar flow hoods and two inverted microscope for cell and tissue culture from Leica. ELISA plate shaker; 96-well plate washer; ultracentrifuges; Environ Shaker from Lab-Line; Centrivap Concentrator from Labconco; Gel Dryer from Bio-Rad; WKL 230 Water Cooler from Brinkman; HM550 Cryostat from Thermofisher; Biological Safety Cabinet from Forma Scientific; Heracell 150i Tri-Gas cell culture incubator from Thermo Scientific; Bio-rad TC10 automatic cell counter; Hugo Sachs Elektronik Easy Cell Extraction System Type 803 for isolation of primary cardiac fibroblasts and myocytes via aortic perfusion of whole hearts from Harvard Apparatus. Odyssey Infrared protein detector system: this system provides a highly sensitive and accurate way to quantify immune-detected proteins on blotting membranes. It also has the capability for simultaneous detection and quantification of multiple proteins. 

The core is equipped with a Leica TCS SP5 Laser Scanning Confocal Microscope with a Multi-photon Laser and is located within the University of Toledo's Advanced Microscopy & Imaging Center.  Specifically, the Leica TCS SP5 is equipped with both conventional and high-speed resonance scanners. The Multi-photon laser is attached to an inverted microscope and allows deeper penetration into tissue with less photo-bleaching/damage. Five lasers plus multi-photon excitations are available, producing the following laser excitation lines:  458, 488, 514, 561, 633, and 710-990nm (from the tunable Ti-Sapphire MP laser).  The system is capable of collecting up to 7 colors simultaneously for quantitative confocal image analysis, 3D reconstruction, FRAP and FRET, animation, stereo imaging, single layer projection, time lapse collection, and co-localization analysis.