Preclinical Imaging Centre (PRIME)

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2 Preclinical Imaging Centre (PRIME) Visiting address PRIME (route 231) Geert Grooteplein EZ Nijmegen The Netherlands Nicole Bakker 31 (024) At the Central Animal Facility (CDL) a state-of-the-art preclinical imaging centre is housed. This Preclinical Imaging Centre (PRIME) is a partnership between the departments of Radiology, Nuclear Medicine, Cell Biology, Rheumatology, CDL, Cognitive Neuroscience and Anatomy of the Radboud University Medical Center Nijmegen. Each department provides dedicated state of-the-art equipment and expertise for small animal imaging and laboratory animal science, including PET/CT, SPECT/CT, MRI, an irradiator, intravital multiphoton, fluorescence/bioluminescence imaging, high frequency ultrasound and behavior facilities. These preclinical non-invasive imaging technologies have emerged as powerful resources to monitor physiological and disease processes in mouse and rat models. Combining the strengths of anatomical and functional imaging modalities with cognition and behavior allows the detection and quantitative determination of morphological, metabolic and functional processes in live animals. Imaging equipment and other hardware available Magnetic resonance imaging (7 and 11.7T ) Radionuclide Imaging (micropet/ct and microspect/ct) Optical Imaging System (IVIS Lumina) Two-photon fluorescence microscopy High frequency ultrasound (Vevo-2100) Irradiator Technical assistance to run the instruments Probe preparation Data processing and analyses Wet lab space Behavior and Cognition Facilities Animal facilities and animal care and treatment

3 Behavior and Cognition Facilities Measurements of behavior and cognition and in combination with brain imaging are indispensable for corroboration of the relation between e.g. cognition and brain metabolism, blood flow, and white matter (connectivity) as measured with MRS, ASL, DTI and rs fcmri. In the Behavior and Cognition Facilities in PRIME, (social) behavior, cognition, sensory-motor integration and blood pressure in neural development, adolescence, ageing and neurodegenerative diseases can be measured in several mice models, in relation to e.g. lifestyle (diet and exercise) or drug compounds. The EthoVision XT (10) tracking software is used for cognitive tests like the open field test, or tests with mazes such as the Morris water maze. This video tracking software tracks and analyses the behavior, cognition, movement, and activity of animals in multiple arenas or mazes. With the Multiple Body Point Module social interaction between animals can be monitored as well. In the available Phenotyper Systems, animals can be observed automatically for 24 hrs a day in their home cages with an infrared camera. Dr. AJ Kiliaan [email protected] Drs. M Wiesmann [email protected] The available equipment and Ethovision software can be used to investigate: normal behaviour (2x open field) learning and memory (2x Morris water maze, Barnes maze) sensory-motor integration (Rotarod) automated observation in home cages (8x phenotypers) tracking and analyzing (social) behavior, cognition, movement of animals in multiple arenas (2x EthoVision XT (10) tracking software) blood pressure (1x tail cuff plethysmography) SR-LAB (startle response system) for prepulse inhibition

4 Fluorescence and Bioluminescence Imaging Optical imaging offers unique possibilities for in vitro and in vivo imaging applications, especially in the context of molecular imaging. With the IVIS Lumina imaging system it is possible to identify disease pathways, determine mechanisms of action, evaluate drug compounds and monitor their effects on disease progression in living animals non-invasively. This imaging system is highly sensitive and can image fluorescent and/or bioluminescent reporters and probes both in vivo and in vitro. This imaging module allows rapid (1-10 minutes) quantitative assessment of fluorecency in 3-5 mice or 2 rats simultaneously. In addition, a vast amount of biological tools (bioware) are commercially available such as transgenic reporter animals, reporter tumor cells, microorganisms, gene reporters, enzymatic cleavable substrates and quantum dots. In-house nearinfra red fluorescent probes are produced for fluorescence imaging. The IVIS Lumina can be used for the following applications: to study the physiological, pathological and molecular processes in real time and longitudinally. for diagnostics, pharmacokinetics, pharmacodynamics, and drug screening in preclinical studies. Dr. Fons van de Loo [email protected] This imaging module is used for: Tumor imaging with fluorescence imaging Wound healing Skin infection models Malaria infection / immunization Disease-inducible promoters for inflammation Rheumatoid arthritis and osteoarthritis models Dr. Edwin Geven [email protected]

5 Magnetic Resonance Imaging An MR system uses a strong magnetic field to align the magnetization of some atoms in the body, and radio frequency fields to systematically alter this alignment for detection purposes. Using the signals of hydrogen in water, MRI routinely provides good contrast between and within soft tissues in the body. This contrast can be varied for optimal visualization (e.g. of diseased tissue). On top of this, MR can be used for bone and (large) vessel imaging and to obtain functional tissue information such as vascular parameters (blood volume, flow), metabolism, oxygenation, cell density and more. Unlike CT or traditional X-rays, MRI uses no ionizing radiation, but stable isotopes or contrast media to track certain processes or visualize tissues and targets. In the PRIME facility two MR systems with horizontal bore magnets are available: one at 7T field strength (Bruker Clinscan) and another at 11.7T field strenght (Bruker Biospec). Both systems have a large range of radiofrequency probes, mostly focusing on mice and rat applications (e.g. for muscle, brain). Both systems have anesthesia and animal monitoring devices. Prof. Dr. Arend Heerschap [email protected] Ing. Andor Veltien [email protected] The scanners can be used in the following applications; Anatomy and morphology Metabolic (spectroscopic) imaging Perfusion and vascular imaging Diffusion and diffusion tensor imaging Targeted contrast MR, e.g. in cellular imaging The applications can be performed quantitative

6 Multi-Photon Microscopy Multi-photon excitation microscopy (MPM) is used for fluorescence imaging of living three-dimensional tissue at subcellular resolution up to one millimeter in depth. The MPM setup in PRIME consists of two Trim Scope-II microscopes (LaVision Biotec, Germany), including three fs-pulsed Titanium: Sapphire lasers (Chameleon Ultra, Coherent) and an optical parametric oscillator (Coherent-APE) to excite samples with two near infrared and one infrared wavelength simultaneously. Up to 7 non-descanned detection channels are available in the backward and forward direction, including two 16- channel time domain fluorescent lifetime detectors for intravital fluorescent lifetime imaging. Infrared MPM reaches up to 10-fold improved excitation of far-red fluorophores (e.g. DsRed, mcherry), two-fold tissue penetration, reduces photo bleaching 4-10 fold and minimizes photo toxicity, compared with conventionally used nearinfrared excitation, thus maximizing sample volume and biocompatibility during intravital microscopy. Prof. Dr. Peter Friedl [email protected] Dr. Gert-Jan Bakker [email protected] Long/term serial and time-lapse imaging of body surfaces and live tissue in window models, with focus on cancer biology and experimental therapy. Endpoint imaging of deeper organs The response of different tissue regions to therapeutic interference, Co registration of functional parameters after application of functional reporter probes. Imaging of fixed tissue samples Two-photon excited laser microsurgery of cells and tissues (3 µm precision) Monitoring of cancer invasion and leukocyte dynamics in thick organotypic 3D cultures 3D image reconstruction, image segmentation and quantification using standard and customized software

7 Multi-Photon / Intrinsic Optical Imaging The combination of intrinsic optical imaging and multiphoton microscopy has many applications in neuroscience, because it allows to measure neural activity and structural changes in the nervous system at multiple scales. With intrinsic optical imaging the haemodynamic response and large-scale neural network activity can be measured in cortical areas at the mm-scale. With two-photon imaging the activity of large scale neural networks of individual neurons can be recorded and structural measurements can be performed up to the scale of individual dendritic spines. The two-photon imaging set-up at PRIME contains a 8000Hz resonance scanner which allows for scanning at a high temporal resolution (512x512 pixels at 30Hz.) The set-ups are designed to be combined with viral injections (e.g. optogenetics), patch-clamp and other neurophysiological techniques, visual and somatosensory stimulation and behavioral tasks on a running ball. The intrinsic optical imaging set-up and two-photon imaging set-up are specifically designed for recording in: Prof. Dr. Richard van Wezel [email protected] Prof. Dr. Tansu Celikel [email protected] Cultured neural networks Organotypic chronic slice cultures Anaesthetized animals Chronic recordings in animals performing behavioral tasks The imaging modules are currently used for: Recording structural changes of dendritic spines In vivo recording of calcium transients in large neural networks Studies on visual, somatosensory and other cortical areas

8 Radionuclide Imaging and CT Radionuclide imaging, including single photon emission computed tomography (SPECT) and positron emission tomography (PET), relies on the tracer principle in which a tracer amount of a radioactive probe is injected into the body to monitor a physiological/ biochemical process. By combining these imaging with an x-ray Computed Tomography (CT), the images acquired with both devices can be acquired sequentially, in the same session. Thus, functional imaging obtained by PET or SPECT, which depicts the spatial distribution of metabolic or biochemical process, can be aligned with anatomic imaging obtained by CT scanning. At PRIME a wide series of Tc-99m, In-111, I-125, I-123-labeled probes are used for SPECT. PET probes labeled with F-18, Ga-68, Zr- 89 and I-124 are produced. Prof. Dr. Otto Boerman [email protected] Dr. Peter Laverman [email protected] The PET/CT and SPECT/CT scanners can be used for the following applications in mice and rats: imaging metabolic/biochemical processes imaging receptor expression imaging gene expression imaging in vivo migration of cells The CT scanner (i.e. without PET or SPECT) can be used for the following applications in mice and rats: High resolution (20-40 µm) focused CT scans Medium resolution ( µm) whole body CT scans Bone density measurements

9 High Frequency Ultrasound Imaging High Frequency Ultrasound Imaging is a technique used to visualize detailed internal structures with unprecedented temporal and spatial resolution. By transmitting high frequency short ultrasound pulses and receiving these pulses reflected by internal tissue structures, images with a resolution down to 20 microns can be obtained at up to 500 frames per second. Ultrasound is safe, does not use ionizing radiation and is easy to use. The ultrasound transducers can be positioned on the region of interest with only using coupling gel. The Visualsonics is equipped with a 20, 40 and 50 MHz transducer. The higher the frequency, the better the resolution, but the lower the imaging depth. Additionally, functional imaging can be performed using Doppler techniques for quantification of flow and dedicated ultrasound contrast agents for tissue perfusion Dr. Chris de Korte [email protected] Cardiac imaging in small animals Organ imaging (e.g. kidney, liver, brain, muscles) in small animals Ex vivo imaging of tissue

10 Central Animal Laboratory The CDL offers all facilities to perform high quality animal related research and education. Housing facilities Next to conventional housing the CDL can offer a variety on specific housing facilities for most laboratory animal species. Surgery facilities The CDL provides the necessary basic infrastructure for surgical research. Full equipped operating theatres, radiology room and area for sterilization of instruments are available for academic research and industry. Facilities for surgical product testing and evaluation and for demonstrations and instructional courses for instance for training of hospital staff are available. Biotechnical support Well trained and competent personnel will help to plan and execute the proposed research, product testing, instructional course or demonstration. Our biotechnical staff can assist you with various technical activities varying from injections of compounds by all recognized routes and collection of tissue for futher analysis up to complex surgical procedures. [email protected] 31(024) SYRCLE SYRCLE is a unique and innovative research and education centre focusing on the continuous improvement of scientific quality and transparency of laboratory animal science and of animal welfare. Its current focus is on systematic reviews. These help to improve the quality of scientific research, the search for, and implementation of, the 3Rs (Refinement, Reduction, Replacement) and patient safety and translational medicine. The 3RRC facilitates the development of guidelines, as well as the execution and education of systematic reviews.