CNY-5

The 5th floor of CNY contains the main CMIR laboratories, Administration, Cellular Imaging Program (CIP), Laboratory for Biooptics and Molecular Imaging (LBMI) and High-Throughput Screening Facility (MLIA).

1. Chemistry Laboratory

The chemistry laboratory is specifically designed for the synthesis of novel imaging pharmaceuticals. These laboratories consists of 3,000 sq. ft. of modern research and production dedicated space equipped with acid resistant benches. This laboratory holds over 3,500 reagents and all necessary glassware and small equipment. In addition to bench space there are three walk in refrigerators for low temperature syntheses, and six Hoodaire airflow fume hoods, equipped with gas, vacuum, and water. The laboratory furthermore contains two modern spectrophotometers (Fluorolog 3, Varian Cary50) and two spectrofluorometers (Hitachi U-3000, Jobin Yuon) all connected to the network. There are four HPLC systems: 1) an analytic HPLC (Varian, ProStar) and 2) three preparative HPLC (Rainin, HPXL, Hitachi 7000) and an automated flash chromatography system (Biorad, Econo). Additional equipment includes a fully automated Rainin PS3 peptide synthesizer, an Advanced Chemtech Apex 396 combinatorial peptide synthesizer, two Buchi 011 Rotavapors, Precision and Thermoline incubators, Gyrotron G2 orbital shaker, pH meters (Orion), several stirrer/hot plates (Corning PC351, 510), several Amicon stirred cells (8010, 8050, 8200, 8400), an oven (Fisher Isotemp 200 series, model 230G) and fluorescence analysis cabinet for thin layer chromatography. A Bruker 120 Minispec pulse NMR spectrometer is available for T1 and T2 measurements. The lab furthermore contains a Branson sonifier 450, a RC5 high-acceleration centrifuge (Sorvall), a Sorvall ultracentrifuge, several Eppendorf microfuges, a submicron dynamic light scattering particle analyzer (Zetasizer 1000HS, Malvern), a lyophilizer (Virtis), a high speed, micro volume concentrater (Savant, SpeedVac AES1010), two high efficiency flow dialysis systems (Amicon, CH2 and A/G Technology, QuixStand), a fluorescence plate reader (Molecular Devises, Spectramax) and water purification system (MilliQ, Millipore).

2. Biochemistry Laboratory

The biochemistry laboratory is designed for the evaluation of pharmaceuticals, their interaction with biological material and for biochemical analysis. This laboratory consists of 1,200 sq. ft. of modern laboratory space equipped with acid resistant benches, appropriate shelving and small equipment (glassware, pipettes etc.) and contains over 1,000 biological reagents. There are several Amicon cells (volume from 10 to 300 mls) for concentrating protein solutions and a set of membranes with cut-off range from 3 to 300 kDa. This laboratory also contains several Bio-Rad mini protein II cells and complete electroblotting systems (Bio-Rad), an osmometer (Osmette A), two Mettler AE 200 balances, and a Pharmacia GE-24 gel electrophoresis unit with an EPS 500/400 power supply, and a Bio-Rad Econo low pressure chromatography system. The lab contains several water baths (Haake A80, Haake F3 and Precision 2), and fraction collectors (Pharmacia) as well as a fume hood and a sterile laboratory hood. A walk in cold room (4°C) and freezer room (-20°C and -70°C) are available. There are several refrigerators/freezers (VWR). A Kodak M35-X-omat film processor is available for processing autoradiographs. The dark room (120 sq. ft.) is located adjacent to the biochemistry lab.

3. Molecular Biology Laboratory

This facility was recently expanded and is equipped for the isolation, purification and production of DNA, cloning, DNA transfection and investigations in gene therapy. The laboratory currently consists of several rooms (2000 sq. ft.) equipped with all necessary equipment. The facility contains equipment for horizontal submarine electrophoresis (Bio-Rad, Owl Scientific), portable laminar flow hoods for bacterial culture, (Fisher, model 785), vacuum blotters, and two incubators/shakers for bacterial culture. Sequi-GenII sequencing cell (Bio-Rad), thermocyclers (MJ Research and Stratagene), an incubator hybridization chamber, Sorvall RC28S centrifuge (Kendro Laboratory Products), Ultracentrifuge L8-55 (Beckman) and three freezers (- 20 oC and - 70 oC, Gibson and Revco). In addition, we have recently acquired a new Molecular Dynamics Phosphor imager SI, an Image Eraser SF, with phosphor and tritium screens, as well as accompanying software. There is a BioMax/BandScanner Scientific Imaging Systems (Kodak) for DNA/RNA and protein gel analysis as well as a new Kodak Digital Science TM Image Station 440CF (IS440CF) system, providing very sensitive and quantitative imaging of chemiluminescence, fluorescence and chromogenic gels and blot analysis.. There is also an Arcus II photoscanner (AGFA) with accompanying software for efficient analysis of radiolabeled samples. The facility also has the necessary softwares for the PCR primer design Amplify 1.2 (Bill Engels) and Primer Premier 4.0 (Bosoft Intl.) and Primer Express (Applied Biosystems); MacVector (Oxford Molecular Group) and Vector NTI Suite (InforMax) for the vector design; Sequencher - for the DNA/RNA sequence analysis and EditView (Perkin-Elmer) for analysis of sequencing results. An ABI Prism 7900HT sequence detection system, for quantitative PCR, is also available in this lab.

4. Radiochemistry Laboratory

All CMIR laboratories are approved for handling of radioactivity (including 111In, 125I, 99mTc, 32P, 33P, 35S, 3H, 59Fe, 153Gd, 51Cr, 210Tl, 14C, 141Ce, 95Nb, 103Ru, 46Sc, and 113Sn). The CMIR has two designated hotlabs. One lab is designated for general radiochemistry and the other for radiochemistry in Molecular Biology. The general radiochemistry lab consists of approximately 200 sq. ft. of lab space and contains a Capintec dosimeter, Skatron semi-automatic cell harvester, Fisher Scientific sonic dismembrator, ROSI reciprocating orbital shaker incubator, Marvel refrigerator, Kenmore freezer, two Forma Scientific water jacketed incubators, as well as all other necessary equipment for a chemistry laboratory. The Molecular Biology hot lab is approximately 125 sq. ft and contains a Hoodaire laminar airflow hood, MJ Research mini-cycler, Stratagene UV stratalinker, Robbins scientific micro hybridization incubator, Fisher Scientific gel dryer, electrophoretic apparatus, as well as all other necessary equipment. A new Wallac 1480 wizard gamma counter and Packard Tri-Carb liquid scintillation analyzer are available for biodistribution studies. An additional dedicated facility of approximately 200 sq. ft. is also available for iodination purposes and is a common laboratory located adjacent to the general lab space.

5. Cell Culture

This facility was established for the culture of tumor cells, normal tissue cultures, production of monoclonal antibodies and cell transfection. The CMIR tissue culture laboratory consists of four adjacent rooms (150 sq. ft. each) that are equipped with a total of five laminar flow hoods (Forma Scientific model 1132), seven dual chamber CO2- incubators (Forma Scientific water jacketed), three microscopes (Nikon TMS), waterbaths (Labline waterbath) and benchtop centrifuges. There are also three Biogard liquid nitrogen storage tanks for storage of cells. In addition there is a new four-color FACSCalibur flow cytometer (Becton Dickinson) and dedicated analysis station running on a Mac G4 computer.

6. Histology and Microscopy Laboratory

This facility was recently installed and consists of a dedicated microscopy room (150 sq. ft.) and additional lab space for tissue sectioning, embedding and staining (15 ft acid resistant bench, fume hood). The facility contains a Reichert-Jung microtome, a cryotome (Leica, International Equipment) and a tissue staining device (Technicon Co). An Axiovert 100TV Zeiss inverted light and fluorescent microscope is available with a water heated microcopy chamber for live cell work. This microscope has attached to it an automated film camera system (MC80 system, Zeiss) and also contains a high resolution, low light level CCD camera (Photometrics) that is attached to a Mac G4 computer. An upright Nikon light microscope with color CCD camera attached is available for immunohistochemistry slides and a further inverted Nikon Eclipse TE2000 equipped for transmitted light and epifluorescence with a second color CCD camera (Insight QE) is available for live cell analysis. Both are connected directly to a Mac G4 computer with designated analysis software.

7. Robotics and High Throughput Screening

The High Throughput Screening lab consists of a Beckman Coulter Biomech 2000 automated liquid handler, a Nikon Eclipse TE 2000S fluorescence microscope, and an associated cell culture area. The liquid handler is equipped with 4 stacker towers for storage of plates and tip boxes, a 6 port wash unit, a gripper tool for moving components, a Brinkman Lauda WKL230 recirculating bath, and assorted liquid handling pipettes. In the near future it will be further functionalized with an integrated plate reader (Molecular Devices, Spectromax) and a filter station; allowing automated high through put chemistry and cell uptake studies. Associated with the High Throughput facility is a fourth 150 square foot cell culture facility, equipped with an additional laminar flow hood (ThermoForma), 4 water jacketed CO2 incubators (ThermoForma), and a Beckman Coulter Z1 Coulter particle counter. For additional cell culture facilities see below.

8. Biomedical Engineering

The main direction of this laboratory is the development of novel optical imaging and and other imaging technologies. Members of the lab have recently completed the development of two Fluorescence Molecular Tomography systems (FMT) with different performance characteristics. They have also constructed a time-resolved spectrometer for measurements of optical properties in diffuse media as well as a highly sensitive planar imaging system for bioluminescence imaging. The lab also supports image processing methodologies.

9. CMIR IT Infrastructure

At the core of the CMIR IT Infrastructure is MIPortal, a high-capacity information platform. It enables the local storage of large data sets, enhances real time availability of diverse data, facilitates multidisciplinary and inter-institutional research, and serves as a platform for long term data storage and subsequent searches, databanks and analysis.
All hardware is enclosed in a single HP computer rack and contains 5 servers (Application Server, PACS Server, Network Attached Storage, Back-End Server, and Staging Server), disk array, tape library, DVD Jukebox (NSM 3000 DVD Jukebox with 225 loaded DVD, 3.5 TB capacity). Storage is provided by a HP StorageWorks NAS 2000s server with 3 TB of raw disk space expandable to 12 TB. A MSL5030 tape library with HP OpenView DataProtector software provides automated backup capability. All components are connected via a local Gigabit network. Image post-processing is performed on two dedicated diagnostic workstations (Siemens Leonardo) and two dual-CPU G5 Macs.
All computers are currently networked via Ethernet to the Internet. In addition to the MIPortal system, the computer network contains the following equipment: SGI Silicon Graphics Octane workstation, over 80 Macintosh computers and over 70 PCs. Three laser printers (HP Laser Jet 4M and 2 4050N), a photo-quality color printer (Kodak ds 8650 PS Color Printer), 4 ink jet color printer as well as AGFA Arcus II scanner are also available. The facility has available all necessary software for bioinformatics and chemoinformatic research.

10. Office Space

The office space consists of eight (250 sq. ft. each) offices, some of which are equipped with a FAX and additional computers. There is a large conference room and a lounge.

11. Animal Farm

The MGH Charlestown Animal Facility has exceptional animal housing facilities and abundant laboratory work space located in the same building as the laboratories. An additional animal farm is located in the MGH Wellman Building. In both facilities, common area space is available for animal surgery, autopsy, and tissue handling

The MGH 8th floor Central Animal Facility is a Viral Antibody Free (VAF) area for rodents, with well established containment procedures, and contains approximately 10,700 sq. ft. of state-of-the-art animal housing space. The area is equipped with a cage/rack washing facility and autoclave. Two 250 sq. ft. limited access animal rooms and attached procedure room for the exclusive housing of immunocompromised rodents located within the 8th floor Central Animal Facility, and will be used for housing all mice. The procedure room to be used for studies under this proposal is equipped with a tissue culture hood, centrifuge, CO2 incubator, refrigerator, microscope and bench space. Animal care in these spaces is under the direction of the Director, Office of Laboratory Animal Resources, and ACLAM certified veterinarian.