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Equipment
| BRC TECHNOLOGY AND EQUIPMENT | ||||||||||||||||||||||||||||
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Technologies developed and available at the BRC focus on the study of the living cell, with a particular emphasis on documenting chemical activity in the spaces adjacent to the external face of the plasma membrane. Go to complete resource list >> |
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ELECTROCHEMISTRY & PHYSIOLOGY
SPINNING DISK CONFOCAL - A Perkin Elmer spinning disk scanning unit (CSU-21) plus Orca CCD (Hamamatsu) is installed on a Zeiss Axiovert 200 inverted microscope with integrated electrochemical and electrochemistry micromanipulators and acquisition hardware. The multi-line argon laser (Melles Griot) accommodates 488, 568 and 647nm laser lines. Epifluorescence is available on this system via an illuminator (EXFO). It is housed on an air suspension table, in a darkened Faraday box, which can be heated to 37oC. 9 RESEARCH & EXPERIMENTAL PLATFORMS which can be configured for self-referencing electrochemistry and standard patch / sharp microelectrode recording for current and voltage clamp. These include headstages and amplifiers from Molecular Devices as well as BRC headstages and controller units. Four platforms also support FLUORESCENCE GLOBAL IMAGING on 3 inverted and 1 upright Zeiss microscope. Micropositioning is performed with many different commercially available fine and coarse, 3-axis positioners including 4 fine controlled Narishige MHW-3 positioners. MICROPIPETTE & MICROSENSOR FABRICATION: The BRC has two Flaming/Brown micropipette pullers (P97; Sutter Instruments) and one laser-based puller (P2000; Sutter Instruments). These are housed in a dedicated clean room. DIELECTROPHORESIS SYSTEM: The wave form generator for developing the dielectrophoretic system is already available at the BRC – indeed it was designed and built there. This system is portable and can operated from any of the experimental platforms in BRC or the MBL Central Microscopy Facility. |
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| IMAGING (also see Electrochemistry & Physiology above) LOW LIGHT: Described above this unique system can be integrated with electrochemistry and electrophysiology. It is housed on an airtable in a darkened Faraday box which can be held at 37oC. SPINNING DISK: A Perkin Elmer spinning disK is installed on a Zeiss inverted scope and integrated with electrochemistry and electrophysiology. It is housed on an airtable in a darkened Faraday box which can be held at 37oC. FLUORESCENCE CALIBRATION - Horiba FluoroMax-3 and MicroMax-384 systems are available for characterizing sample fluorescence properties. << Top |
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| MOLECULAR BIOLOGY TISSUE CULTURE: The BRC maintains a culture room with the necessary equipment for mammalian cell culture. This includes: • Two laminar flow sterile hoods, one with a low front glass for common culturing and the other with an open front for use with a dissection microscope. • Two 5% CO2 37o C incubators for maintaining cells, and a liquid N2 dewar and -80o C freezer for storing cell lines and reagents for long periods of time. RNA & DNA ISOLATION & ANALYSIS, restriction analysis, protein isolation and analysis: western blot & immunoblot cloning TRANSFECTION of clonal and primary cells using: • Adenoviral/lentiviral vectors used with cardiac and neuronal cells respectively. Vectors: gene of interest (GOI) under the CMV or RIP (=rat insulin promoter) • Electroporator (Micropulser; BIORAD) used primarily for bacterial transformation. • Magnetofection (OZ BioSciences) optimized for clonal and neuronal cells. • Ca2+ phosphate transfection optimized for neuronal cells. CLONING • Gradient Thermocycler (BioRad, Hercules, C.A.) • Electrophoresis systems (BioRad, Hercules, C.A.) • Bio Doc gel imaging system (UVP, Upland, CA) • ND-1000 Spectrophotometer (NanoDrop, Thermo Fisher Scientific, Wilmington, DE); quantification of DNA and RNA • Bacterial culture 37o C incubator and shaker • Transfection with Lipofectamine 2000 (Invitrogen), magnetofection (OZ Biosciences, Marseille, France) << Top |
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MBL CENTRAL MICROSCOPY FACILITY This is a fee for service facility adjacent to the BRC laboratories in the Lillie Building. The BRC has access to: • Two Zeiss LSM-510 NLO with META detector laser scanning confocal systems. One is configured on an inverted Axiovert 200 M microscope the other on an upright Axioscop 2FS. The Zeiss 510 LSM has three independent simultaneous confocal channels for reflection and fluorescence and an extra channel for transmitted light. Both systems are capable of standard and 2 photon excitation using the Coherent Chameleon lasers. These highly automated systems create 8 or 12-bit 2048x2048 digital images using Pentium PC based user interfaces. The Zeiss 510 LSM is configured on an inverted microscope with a variety of objectives available and a complete incubator chamber system. Both the inverted and upright confocal scopes allow fluorescence spectral analysis using diffraction grating, with 10nm spectral channel width. They also possess a 32 channel anode array detector system which has a pinhole of variable size for each detector. • Both TEM and SEM are available in the same facility. The TEM is a Zeiss 10CA 20-100KV. The SEM is a JEOL JSM-840 1- 35KV. • A new SEM is to be purchased which will have a field emission gun source (FE) and will include high vacuum and variable pressure modes. It will have a new elemental dispersive spectrometer system (EDS) along with electron backscatter diffraction system (EBSD). It will also have a cryostage and automated standard stage. Full EM preparation and microtome facilities are available at the same location. COMPLETE RESOURCE LIST >> |
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