BIOIMAGING (Under Construction)



Bioimaging – James Keen, PhD, Kimmel Cancer Center (KCC), TJU.
The goal of the KCC Bioimaging Shared Resource is to provide detailed light microscopic image analysis capabilities in basic and medical science research programs. The Bioimaging Shared Resource operates with a well-trained technician and a faculty supervisor, Dr. James Keen. The Bioimaging Shared Resource provides investigators with reliable instrumentation operated with a high degree of technical expertise, and relief from the obligation for substantial capital investment for equipment.
Confocal Laser Microscopy. The Bioimaging Shared Resource is equipped with a state-of-the-art Zeiss LSM 510 META Confocal Laser Scanning Microscope System interfaced to a Zeiss Axiovert 200M inverted microscope equipped with 5 objectives ranging from 10x to 63x. The confocal uses multiple lasers with excitation lines at 405 nm, 458 nm, 477 nm, 488 nm, 514 nm, 543 nm, and 633 nm. The system's 3 PMT detectors provide the ability to simultaneously image up to 8 fluorophores with emission between 417 nm and 730 nm. Widefield microscopy: This resource is equipped with a widefield epifluorescence imaging system specifically designed for high quality acquisition of living cell images. This “Live Cell System” is built around a Zeiss Axiovert 200M inverted microscope equipped with 5 objectives ranging from 10x to 100x that can be used for both epifluorescence and brightfield microscopy. Microinjection: This system is set up on a Nikon Eclipse TE-2000-S inverted microscope with an attached Eppendorf Micromanipulator 5171 and Microinjector 5242. The instrument allows adjustment of injection angle for injection of cultured cells or C. elegans.



Bioimaging – Joseph Rabinowitz, PhD, Center for Translational Medicine, TJU.
Within the Center for Translational Medicine (CTM) at TJU, Dr. Rabinowitz has technology available through his laboratory to image light emission and fluorescence in live animals, critical to stem cell and gene therapy research being done in the CTM. The in vivo bioluminescent system is the Xenogen Series (now Caliper) IVIS 100 Imaging System. It is sensitive and has a large field of view. The focusing stage is adjustable from 10-25 cm resulting in a field of view large enough for 2 rats or 5 mice or small enough for only 1 mouse. The system features a 25 mm (1.0 inch) square back-thinned, back-illuminated CCD, which is cryogenically cooled to greater than –100°C via a closed cycle refrigeration system to minimize electronic background and maximize sensitivity. The systems animal handling features include gas anesthesia within the chamber and in a specialized holding compartment. The chamber has a heated shelf to maintain the comfort of the animals during imaging. The Living Image software is easy to use and to transfer files for presentations.



Small animal imaging – Matthew Thakur, PhD, KCC, TJU.
The Small Animal Imaging Shared Resource provides state-of-the-art in vivo molecular imaging capabilities. It was established under the direction of Dr. Matthew Thakur in April 2004 and is supported, to date, by TJU and the Department of Radiology. This resource provides non-invasive small animal imaging, with PET, SPECT, CT, and ultrasound. These modalities offer innovative basic, as well as pre-clinical translational, research capabilities for studying pathophysiology and genesis of cancer. The facility is supported by 3 AALAC accredited animal houses and 2 veterinarian staff members. The imaging facility is housed in 1018 Bluemle Life Science Building. It is strategically located adjacent to the animal housing where a majority of the Cancer Center laboratory animals reside and in the building where a majority of the investigators perform cancer research. The 250 ft2 imaging facility currently houses the Philips Mosaic PET scanner, the Imtek microCAT II CT scanner and a host of support equipment.
The Mosaic small animal PET scanner (Philips Medical Systems) uses 2x2x10 mm3 GSO (gadolinium oxyorthosilicate) crystals coupled to 19 mm diameter photomultiplier tubes (PMT) via a continuous slotted light guide. The detectors are arranged to produce a transaxial field of view (FOV) of 128 mm and an axial FOV of 120 mm. The absolute coincidence sensitivity is 1.3% for a point source and the transverse resolution is 2.2 mm at full-width half- maximum (FWHM). The ImTek microCAT II small animal CT scanner has 2 main roles: as an ultra-high resolution system for anatomical imaging down to 50 μm, and to generate anatomical reference images for correlation with the functional PET and SPECT data. Radionuclide images typically lack sufficient anatomical detail to permit delineation of organs, thus requiring a co-registered reference image from a modality such as CT. Supporting equipment in the imaging facility includes laminar flow hood, injection stages, infusion pump, micro centrifuge, dose calibrator, scintillation counter, survey meter, shielded assay area, small animal bio-monitoring system, image processing and analysis computers and software.



Small animal imaging and models of human disease – Walter Koch, PhD, Center for Translational Medicine, TJU.
The Center for Translational Medicine (CTM) at TJU has invested in operation of a Surgical and Physiological Core Unit headed by Dr. Erhe Gao and currently staffed by 3 professionals. This Core unit can measure all cardiovascular parameters in vivo including EKG and blood pressure using telemetry, invasive hemodynamics including cardiac pressure-volume loops and finally in vivo echocardiography to measure cardiac function and dimensions. The CTM has the only Visualsonics system at TJU to deliver small animal ultrasound and although currently dedicated to cardiovascular studies can be outfitted for other applications as well. In addition, this Core facility has a c-ARM fluoroscope and surgical table to support large-animal pre-clinical studies where cardiac imaging is needed. The disease models currently targeted in this Core are myocardial infarction, myocardial ischemic-reperfusion injury, pressure-overload cardiac hypertrophy, kidney injury to induce renal-mediated hypertension



Bioimaging – Kirk Czymmek, PhD, DBI, UD.
The Bioimaging Center at the Delaware Biotechnology Institute is a multi-user facility that provides state-of-the-art microscopic imaging instrumentation and technology. The center is open to researchers and collaborators on a fee-for-service basis; outside industrial users are accommodated when scheduling permits. Access to the center is available through a webbased reservation system. A professional staff of 5 experts under the leadership of Dr. Kirk Czymmek is available for project consultations and provides regular user training. Instruments include: Transmission Electron Microscope (TEM) – Zeiss CEM 902 with a Mega View II digital camera from Soft Imaging Systems; Field Emission Scanning Electron Microscope (FE-SEM)– Hitachi S4700 with a Gatan Alto 2500 Cryotransfer System and an Oxford INCA Energy (EDS) System; The FE-SEM has collaborative capabilities via the Internet. Multiphoton/Confocal Microscope – Zeiss LSM 510 NLO with a PECON Environmental Incubation System; High Speed Spectral Confocal – Zeiss LSM5 DUO, with META and Live Scanhead; Atomic Force Microscope (AFM) – Veeco Nanoscope IIIA with Phase capabilities; Laser Capture Microdissection Microscope (LCM) with a PALM MicroBeam Laser System; Zeiss Axioskop2 and Zeiss M2BIO microscopes using either the Zeiss AxioCam or Hamamatsu Orca-ER digital camera; Microtomy Tools include the Reichert-Jung Ultracut E microtome and the LKB 7800 Knifemaker. Image Analysis and Enhancement workstations: 2 Dell computer systems that have an array of image processing and analysis software, a Fujifilm Pictography 4000 Printer with publication quality print capability, In addition, the center has a facility for specimen preparation and a darkroom with a Kodak X-Omat.



Bioimaging – Bruce Boman, MD, PhD, CCHS.
The Center for Translational Cancer Research (CTCR) currently has a Zeiss motorized AxioObserver Z1 equipped for brightfield, phase contrast, varel contrast, and epi-fluorescence, with both color and monochrome digital cameras, X-Y scanning stage, anti-vibration platform, 10x, 20x, 40x and 100x objectives, and Apotome confocal imaging device with a highperformance PC, a flat-panel monitor, complete Axiovision software solution, and an Axiocam MRm digital camera. The system will be upgraded to a complete Zeiss LSM 710 laser scanning confocal microscope in 2009.



Imaging – Jeffery Twiss, MD, PhD, Nemours/AIDHC.
The Imaging Shared Resource Laboratory provides access to the LEICA DMIRE2 Confocal Microscope System. Training and troubleshooting support is available for microscopy and image processing software.