AMRIS Admin Part of the McKnight Brain Institute Operates as a core facility to UF investigators as well as supports the NHMFL external user program Under federal CAS we set fees to break even--we currently do not include amortization of MRI/S systems Salaries: $560,000 + Expenses: $400,000 = $960,000 budget NHMFL support (2009): $350,000 (direct); UF faculty pay their own fees from other grants and cover the rest of the budget $100k of NHMFL money budgeted to equipment/upgrades and $250k budgeted to instrument time and consulting fees for the external user program and pilot studies Currently we budget $5,000 per external user or pilot project NHMFL provides additional $200,000 to fund three development cores headed by Art Edison, Steve Blackband, and Glenn Walter UF provides matching support for development cores Starting in 2010 NHMFL will provide 0.5 FTE for Barbara Beck (RF engineer)

3T Operator: Joy Kidder 3T Engineer: Dave Peterson RF Engineers: Barbara Beck, Dave Peterson NMR applications: Jim Rocca Spectrometer maintenance and scheduling: Dan Plant Spectrometer maintenance: Kelly Jenkins Animal Imaging applications: Huadong Zeng will be starting December 1 st *Pulse programmer *Imaging data processing * Need to build up AMRIS/NHMFL Technical Staff All of these staff members are partially supported by NHMFL funds through hourly fees, either as consultants or through instrument fees.

AMRIS Instrumentation Joanna Long, Director; Glenn Walter, Associate Director 128 MHz, 3.0 Tesla, Philips whole body MRI/S of humans/large animals 200 MHz, 4.7 Tesla, 33 cm horizontal bore MRI/S of live animals (cats, rabbits, rats, mice, etc.) 473 MHz, 11.1 Tesla, 40 cm horizontal bore MRI/S of live animals (primates, cats, rabbits, rats, mice, etc.) 500 MHz, 11.7 Tesla, 5.2 cm vertical bore Solution/solid state NMR spectroscopy of biomolecules 2x 600 MHz, 14.1 Tesla, 5.2 cm vertical bore Solution/solid state NMR spectroscopy of biomolecules & imaging 1-mm HTS cryoprobe 5-mm commercial cryoprobe MRI/S of superfused cells/tissues 750 MHz, 17.6 Tesla, 8.9 cm vertical bore MRI/S of superfused cells/tissues & of live animals (e.g., mice) Solution/solid state NMR spectroscopy of biomolecules

All instruments have user fees. We must cover all costs without making a profit. Current fees are posted on the AMRIS web site: Most “routine” service jobs can be handled by AMRIS staff New development or very sophisticated applications are done in collaboration with expert faculty groups We budget for approximately 40 external user projects and 10 pilot studies Using AMRIS

AMRIS Mission »» Multi-Frequency RF Probe Development »» Large volume coils »» HTS Probe Development »» Coils Tailored for Unique Capabilities »» High gradients for Diffusion measurement »» MAS Probe Development »» Pulse Sequence Development »» Animal imaging (mouse  elephant) »» Mixture Analysis »» Experimental Protocols User Driven Unique Technology Development for High Field NMR Spectroscopy and Imaging User Driven Development of Unique Applications for High Field MRI/S »» A menagerie of animals (juvenile pig, dog, ibis, rat, mouse, fixed samples) »» Biophysics and Structural Biology »» Chemical Ecology and Metabolomics »» Natural Products »» Diffusion on a nanometer to micron scale »» Low gamma detected MRI/S

AMRIS Facility Highlights ’09 Hired a PhD level Staff Scientist for Animal Imaging – MBI and UF will provide 40% support first three years; other 60% from cost savings measures and current demand Huadong Zeng will start December 1 st New 11.1 T console – SIG from NIH with match from UF and NHMFL; 4 transmit/16 receive; new gradients with integrated shims; new pump station Will arrive in June 2010 Demo 4.7 T system – Funds from NHMFL ARRA leveraged with 11.1 T console purchase; 2006 Demo system from Varian applications; 5 transmit/4 receive; multi-mouse coil; 1996 actively shielded magnet Will be installed in November/December 2009

1-mm HTS Probe 1.Brey, W. W., Edison, A. S., Nast, R. E., Rocca, J. R., Saha, S., and Withers, R. S. (2006) Design, construction, and validation of a 1-mm triple-resonance high-temperature-superconducting probe for NMR, J Magn Reson 179, Dossey, A. T., Walse, S. S., Rocca, J. R., and Edison, A. S. (2006) Single Insect NMR: A New Tool to Probe Chemical Biodiversity, ACS Chemical Biology 1, Dossey, A. T., Walse, S. S., Conle, O. V., and Edison, A. S. (2007) Parectadial: A Novel Monoterpenoid from the Defensive Spray of Parectatosoma mocquerysi Finot 1897, J. Nat. Prod. 70, Zhang, F., Dossey, A. T., Zachariah, C., Edison, A. S., and Bruschweiler, R. (2007) Strategy for automated analysis of dynamic metabolic mixtures by NMR. Application to an insect venom, Analytical Chemistry 79, Srinivasan, J., Kaplan, F., Ajredini, R., Zachariah, C., Alborn, H. T., Teal, P. E. A., Malik, R. U., Edison, A. S., Sternberg, P. W., and Schroeder, F. C. (2008) A blend of small molecules regulates both mating and development in Caenorhabditis elegans, Nature 454, Dossey, A. T., Walse, S. S., and Edison, A. S. (2008) Developmental and geographical variation in the chemical defense of the walkingstick insect Anisomorpha buprestoides, Journal of Chemical Ecology 34, Matthew, S., Schupp, P. J., and Luesch, H. (2008) Apratoxin E, a cytotoxic peptolide from a guamanian collection of the marine cyanobacterium Lyngbya bouillonii, J Nat Prod 71, Kwan, J. C., Rocca, J. R., Abboud, K. A., Paul, V. J., and Luesch, H. (2008) Total structure determination of grassypeptolide, a new marine cyanobacterial cytotoxin, Org Lett 10, Walse, S. S., Lu, F., and Teal, P. E. (2008) Glucosylated suspensosides, water-soluble pheromone conjugates from the oral secretions of male Anastrepha suspensa, J Nat Prod 71, Wang, B., Dossey, A. T., Walse, S. S., Edison, A. S., and Merz, K. M., Jr. (2009) Relative configuration of natural products using NMR chemical shifts, J Nat Prod 72, Dalisay, D. S., Rogers, E. W., Edison, A. S., and Molinski, T. F. (2009) Structure elucidation at the nanomole scale. 1. Trisoxazole macrolides and thiazole-containing cyclic peptides from the nudibranch Hexabranchus sanguineus, J Nat Prod 72, Kaplan, F., Badri, D. V., Zachariah, C., Ajredini, R., Sandoval, F. J., Roje, S., Levine, L. H., Zhang, F., Robinette, S. L., Alborn, H. T., Zhao, W., Stadler, M., Nimalendran, R., Dossey, A. T., Bruschweiler, R., Vivanco, J. M., and Edison, A. S. (2009) Bacterial Attraction and Quorum Sensing Inhibition in Caenorhabditis elegans Exudates, J Chem Ecol. 13.Dossey, A. T., Gottardo, M., Whitaker, J., Roush, W. R., and Edison, A. S. (2009) Chocolate Odorants Sprayed from the Chemical Defense Glands of the Leaf Insect Phyllium westwoodi, J Chemical Ecology In Press Highest mass sensitivity Prototype for Bruker 1.7-mm Several external users—demand for time is high New developments ******************************* “The maglab's 1-mm NMR probe was the key technology that enabled us to identify the mating pheromone in C. elegans. I consider this the most important discovery from my lab in the past decade.” Paul Sternberg Thomas Hunt Morgan Professor of Biology and HHMI, Caltech ******************************* We are working on “the settlement cue for Phestilla nudibranchs that eminates from coral. We have worked on this project for over 10 years and have made more progress in the last two years than we ever did before that. This is due to the ability to get NMR data in the 1-mm probe on tiny amounts of highly bioactive material.” Dr. Valerie Paul, Head Scientist, Smithsonian Marine Station *******************************

Low-E MAS Probe Now available for external users

Optical and magnetic resonance imaging track labeled stem cells within the bone marrow of mice following femoral arterial delivery of bone marrow stem cells. The bone chamber window allows for the optical tracking of transplanted cells prior to high resolution MRI and does not interfere with MRI signal acquisition or have a negative impact on the image quality of MR images. Tibia Window w/ MRI micro-coil for Confocal and MRI Imaging Ultrahigh resolution in vivo MR images obtained following the arterial delivery of labeled bone marrow cells. Images are reconstructed at a resolution of 21x21x38 μm 3 and were acquired in 90 minutes. Dashed red circles indicate areas of image contrast attributed to single cells in both the muscle and the bone marrow. The dashed green circle highlights an area with accumulation of labeled cells. The green legend (line) in A is 150 μm. Stem cell tracking in live mice at 750 MHz G. A. Walter, E. W. Scott, and N.E. Bengtsson, UF; S. Santra; University of Central Florida

900MHz Imaging Coils Single Tune Volume Coils Rat Brain ImagesSodium Images Prototypes Module OD=55mm ID=35mm Length=55mm

750MHz Imaging Coils Double Tune Volume Coils 750 Prototype OD=38mm ID=20mm Length=25mm ProtonSodium ST DT

MRI Microscopy of Mammalian Neurons Steve Blackband Development of Selective Wirelessly-Adjustable Multiple- Frequency Probe (SWAMP) coils Tom Mareci Chemical diversity and geographial variation in insect defensive sprays Aaron Dossey Presentations today

SystemFieldBoreApplicationsAcquiredUpgradedFunding source 4.7T 33 cmAnimal MRI/S , 2009[1][1] UF, NHMFL T5.4 cmNMR, microimaging NHMFL, UF, DoD T5.4 cmNMR1999DoD T8.9 cmNMR, animal MRI/S DoD, State of Florida 11.1T 40 cmAnimal MRI/S DoD, NIH, UF, NHMFL 3T 90 cmHuman,animal MRI/S UF, NHMFL, US Army c T5.4 cmcryocooled NMR2005State of Florida T5.4 cmNMRPendingNIH, UF, NHMFL 7T 31 cmAnimal MRI/SPendingNIH, UF, NHMFL T5.4 cmNMRPendingNIH, UF, NHMFL T31 cmHTS NMRDiscussion2010NIH, Chemistry [1][1] The 4.7 magnet as well as the console will be replaced in Moving forward

Moving Forward - Continuing Facility Enhancement in Gainesville - setting priorities - May ‘09 report to GSB Emergency Needs (list one item) Support for Barbara Beck Top Priority Needs (list three items to maintain existing user program without growth) Additional funds for 11T console upgrade Additional Imaging probe bodies at 600 & 750 Animal monitoring equipment/probe heater Middle Priority Needs (list three items - necessary growth to maintain premier position) Support for imaging scholar/scientist (Receiving from UF/MBI) Console replacements & 14.1 T magnet Animal handler Strategic Planning (list five items - visionary growth in user program or strategic planning) DNP/faculty hire in the area of polarization enhancement MHz NMR system Faculty hire in the area of RF transmission Horizontal, short bore 21T, cm bore magnet 9.4 T, 30 cm bore high-throughput imaging system We would like to have your advice on these or other priorities

Recently Funded or Pending Major Projects Funded –New NIH 1.5-mm HTS probe grant to optimize 1 H and 13 C S/N: Edison, Brey, Larbalestier, Varian. –11 T console upgrade: NIH SIG—requisition went in this week should arrive in June –NHMFL increase from ARRA funds ($200k equipment) will allow us to upgrade 4.7 T with a 3 year old demo system and actively shielded magnet—will be installed in November/December Pending –7T/30 cm animal imager proposal: NIH ARRA HEI –700 MHz NMR for proteins proposal: NIH ARRA HEI –AMRIS facility HVAC and layout renovation NIH ARRA Limited Competition: Core Facility Renovation, Repair, and Improvement (G20) –Helium recovery proposal with UF Physics, High B/T: NSF Academic Research Infrastructure Program: Recovery and Reinvestment (ARI-R 2 ) –950 MHz and 36T console projects previously described

Renovation of the AMRIS Facility Current facility is 12 years old and at capacity. Any new instruments could not currently be housed in the AMRIS wing Specific Aim 1: Reconfigure internal walls within the AMRIS wing to allow addition of new instruments. Specific Aim 2: Renovate HVAC support and cryogen delivery within the AMRIS facility Air handling systems have also improved since the construction of the AMRIS wing. Specific Aim 3: Provide centralized animal staging for imaging and spectroscopy studies

Mechanical 16' Animal Handling 14.1T/54 c14.1T/ T/ T/ T/ T/330 Limited Animal Handling X AMRIS wing

Animal Handling 7T/ T/ T/54 Liquid N 2 Mechanical 14.1T/54 c14.1T/ T/ T/ T/ T/330 AMRIS wing

AMRIS plans moving forward Maintain existing overall general external user program at about the current levels (slight increase). Continued development under 3 technical cores with faculty leaders to better grow unique areas of strength. Build the RF core strengths and continue integrating with Tally RF group. Priorities: –Large volume coils –Multi-nuclear coils –Better 1 H coils/probes for imaging at 750 MHz –Better live animal capabilities

RF Lab Small Animal Imaging Human Imaging

Quadrature Coils for Mouse Imaging Hoult, et.al. MRM 1: (1984) √2 Gain in Signal-to-Noise Ratio Factor of 2 Reduction in Power B1 B A ω 0 M UCGP – Sam Grant, PI Wang, et.al. MRM 48: (2002) Spatial dependence of field magnitude and circular polarization at high freq

Quadrature Coils for Mouse Imaging 25 mm CST Microwave Studio

Preamplifier for Arrays Bottomly, et al, Magn Reson Med, 1990; 16: