SC COBRE in Oxidants, Redox Balance, and Stress Signaling
Cell and Molecular Imaging Core
The COBRE Imaging Core houses five major microscope systems: 1) a Zeiss LSM 880 NLO multiphoton/confocal system with Airyscan super-resolution capability equipped with a Coherent Chameleon tunable femtosecond Ti-Sapphire laser and Quasar spectral detection, 2) a BD Biosciences CARV II real-time spinning disk confocal microscope equipped with a Photometrics Cascade 1K Digital camera and IPLab image processing and analysis software, 3) a Zeiss LSM 510 laser scanning confocal microscope with META spectral detection, 4) an Olympus Fluoview FV1200 intravital multiphoton microscope equipped with a Spectra-Physics Mai Tail tunable Ti-Sapphire laser, and 5) an Olympus Fluoview FV10i laser scanning desktop microscope. The Zeiss and the Olympus microscope systems are equipped with environmental chambers for temperature and gas phase control to allow high resolution non-destructive 3-dimensional imaging of living cells, tissues, small animal organs and organisms. The Olympus Fv10i system has hypoxia control as well. Imaging workstations running Metamorph and other software provide offline image processing and analysis. Major applications of this instrumentation include 1) high resolution live cell imaging of parameter-sensitive fluorophores to monitor ions, electrical potentials, oxygen and nitrogen radical generation, pyridine nucleotide reduction, mitochondrial and plasmalemmal membrane permeability, cell viability (apoptosis and necrosis), fluorescent protein labeling and other parameters; 2) high resolution imaging of tissue sections for immunocytochemistry and green fluorescent protein distribution; 3) fluorescence resonance energy transfer (FRET) to characterize and quantify interactions between specific molecules; and 4) intravital microscopy to monitor microcirculation, leukocyte margination, mitochondrial polarization and permeability, radical generation, gene expression and other factors in living animals. Ancillary equipment includes tissue culture hoods and incubators for specimen preparation; and fluorescence and absorbance plate readers for parallel measurements in cultured cells grown on multi-well plates. Imaging core services will promote the success of the individual COBRE projects and also provide training and assistance to junior investigators studying oxidative stress and stress signaling related to the overall theme of this COBRE. Currently the Imaging Core is supporting 1. Dr. Gaving Wang’s project investigating how cancer stem cells escape anticancer therapy-induced oxidative stress; 2. Dr. Nathan Dolloff’s project of redox regulation in proteasome inhibitor resistant multiple myeloma; 3) Dr. Heather Boger’s study of vagus nerve stimulation for the treatment of Parkinson’s disease and 4) Dr. Peggy Angel’s systems-based analysis of redox activity in aortic valve stenosis. The Core works closely and provides continuous support for graduated COBRE investigators: 1) Dr. Sherine Chan’s project on novel treatments for epilepsy; 2) Dr. Jennifer Isaacs’ study on the role of extracellular Hsp90 in tumor metastasis in prostate cancer; and 3) Dr. Eduardo Maldonado’s project of regulation by voltage gated anion channels of Warburg metabolism in cancer.
John J. Lemasters, M.D., Ph.D.
Professor of Drug Discovery & Biomedical Sciences
GlaxoSmithKline Distinguished Endowed Chair in Advanced Cellular Technologies
Monika Beck Gooz, M.D., Ph.D.
Research Associate Professor of Drug Discovery & Biomedical Sciences