The facilities and resources at the University of Arizona (UArizona) College of Medicine – Tucson (COM-T) are excellent, and the scientific community is vibrant, collaborative, and engaged.

The Sadayappan Laboratory is located on the 3rd floor of the Medical Research Building (MRB) on the UArizona Health Campus in the Department of Cellular and Molecular Medicine. The lab comprises three interconnected open modules with dedicated side rooms for cell culture and microscopic analyses, in vitro functional studies, including cardiomyocyte isolation and skinned myocyte function. The wet benches are equipped with propane and vacuum lines. The laboratory is fully outfitted with state-of-the-art equipment necessary for biochemistry and molecular biology, including rigs for various biochemical and molecular experiments, water baths, an internally calibrated micro-analytical balance, pH meters, and hot-stir plates. All experiments follow safety measures as established by NIH policies and the Institutional Biosafety Committee of the COM-T.

The lab also has access to core facilities at Sarver Heart Center, Cancer Center (NCI Designated), and other basic departments and centers for additional support on complex experiments.

Facilities located in the Sadayappan Lab:

» Calcium and Contractility in Isolated Myocytes: The lab studies the heart at the cellular level using isolated cardiac muscle cells or cardiomyocytes. The IonOptix system allows us to investigate cellular dimensioning and intracellular calcium as important measures of the excitation-contraction coupling.

» Muscle Mechanical Studies: The lab has the latest instruments to measure force development, tension cost, ATPase activity and myofilament calcium sensitivity in skinned cardiac (1) papillary muscles and (2) myocytes (Aurora Scientific Inc).

» Intact Skeletal Muscle Functional Assay: Complete set of Dual-Mode Muscle Lever system (Aurora Scientific Inc) is equipped in our lab to test isometric and isotonic functional capacities of intact skeletal muscle in vivo, in situ, and in vitro.

» Dissecting Microscopes: Numerous Nikon (SMZ-10) and Olympus (BX60) dissecting microscopes are available in the lab for dissecting cardiac muscles for biophysical experiments, as well as taking pictures of TTC-stained heart sections and whole organs post-ischemia- reperfusion injury.

» Fluorescent Microscopes: The lab has an Olympus IX73 fluorescent microscope with a camera attachment to aid in immunohistochemistry documentation.

» In vitro Motility Assay: The lab has access to Nikon A1R LUN-V inverted confocal fluorescent microscope and the reagents required to measure the motility of actin filaments as a function of acto-myosin interaction, along with the rotation of fluorescent myosin rods as a function of mechanical stability in myosin coiled coil.

» Adult Cardiomyocyte Preparation: A Langendorff preparation unit (Radnoti, Monrovia, PA) is available to perfuse the adult mouse hearts and isolate adult cardiomyocytes.

» C02 Incubator and Culture Hoods: The PI has two separate BSL 2-level cell culture facilities. Each room has a biosafety cabinet, C02 incubators, 37˚C bead bath, refrigerator, and inverted microscopes.

Shared facilities available to the Sadayappan lab:

» UArizona Animal Facilities: The University of Arizona has a well-established Research Animal Resources facility, which is managed by professional veterinarians along with well- trained technicians in animal care. Veterinary care is provided by multiple laboratory animal veterinarians and veterinary technicians in University Animal Care, the centralized resource responsible for supervision of veterinary care.

» The UArizona Phenotyping Core: The UA Phenotyping Core offers rodent phenotyping in vivo physiology studies, surgical models, imaging techniques, and functional physiological assessments including echocardiography and electrocardiogram (Visualsonics Vevo F2). The core is staffed by experienced small animal surgeons. Additional phenotyping core information can be found at https://phenotyping.ahsc.arizona.edu.

» Other pertinent shared resources: Induced Pluripotent Stem Cell Core in the Department of CMM; UA Genetics Core (BIO5, DNA extraction, sequencing, fragment analysis, RT-PCR); Proteomics Facility (mass spectrometry analysis of protein molecular weight, sequence, and modification by probes; also, synthesis of peptides and oligonucleotides, sequencing of peptides and DNA); Transmission Electron Microscope Facility hosted by CMM and the Arizona Research Laboratories Biotechnology Imaging Facilities; Statistics Consulting Lab (BIO5 statistical expertise); UA Genetics Core and Genomics Shared Service (GEMM Core, for designing, producing, genotyping, re-deriving, importing, and cryo-preserving genetically modified transgenic and gene-targeted mice).

» Nikon Center of Excellence (CoE) at the UArizona Cancer Center: The CoE is established after the acquisition of several multimodal Nikon microscopy platforms. The direct and immediate benefits of the CoE model to UA include cutting-edge image analysis and priority access to Nikon’s technical expertise for service and customized development of experimental pipelines.

» The UArizona Research Data Center: The UA Research Data Center offers free high performing computing (HPC) resources in a state-of-the-art facility that hosts large computer clusters. UA HPC systems are available to all university faculty, staff, undergraduate and graduate students, and postdocs at no cost.

» The UArizona Viral Production Core: The UA Viral Production Core facility offers services for generating high-titer AAV, Adenoviral, and Lentiviral particles, providing complete cloning services.

» Proteomics Facility: Mass spectrometers are readily available including LTQ Orbitrap, LTQ, QTRAP, Q-TOF, and Voyager DE-STR (For analysis of site-specific phosphorylation and spin labeling); Molecular interactions: Biacore T100 Biosensor (small molecular kinetics, affinity constants, concentration, binding specificity, and thermodynamics); Electrophoresis (1D and 2D SDS-PAGE, for protein and phosphorylation gels); Image analysis and spot picking (ProPicII, ProGest, ProMS, PE Multiprobe; Chromatography: Beckman HPLC, FPLC; lyophilizer, speedvac.

» Functional Genomics Core: High-throughput cherry-picking and plate refactoring, dilution and normalization, and automation: two Dual arm Biomek FX liquid handling automation systems.

For more information, please visit https://research.arizona.edu/facilities

For queries and opportunities, Please Contact :

Sakthivel Sadayappan, PhD, MBA, FAHA
Professor and Head
Department of Cellular & Molecular Medicine
Associate Director, Sarver Heart Center
Czarina M. & Humberto S. Lopez Chair for Excellence in Cardiovascular Research
University of Arizona College of Medicine
Tucson, AZ 85724-5217, USA

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