The Soberman lab consists of 1,500 contiguous square feet of bench space within the Nephrology Division, in the MGH East Research Building. The lab contains a large cell culture facility with three biosafety hoods and four incubators. Of the four incubators, one is dedicated to SILAC (stable isotope labeling by amino acids in cell culture) for determination of proteins in macromolecular complexes by mass spectrometry. There is a dedicated room for use of radioisotopes. A separate area with two additional large benches is dedicated to high performance liquid chromatography and protein electrophoresis. The lab also contains a molecular imaging suite for the study of protein interactions and cell biology. In addition, members of the Soberman lab have access to all shared facilities of the Nephrology Division and other resources described in detail below.
All desks have high-speed Internet access, with connections to all the core facilities and journals.
All members of the lab have state-of-the-art Macs. The lab has two computing systems. One is a wireless Network with back-up hard disks, scanners and printers that integrates the wet lab space, culture facilities, the fellow’s office, and my office. In parallel, all major instrumentation within the lab is connected to a remote server by 1Gb internet ports. The server is connected to 3 workstations in a separate room within the lab’s imaging suite. Each desktop computer within this room is capable of full offline data analysis. All of the lab members and collaborators have their own server space.
• Molecular Imaging Suite
Fluorescence Lifetime Imaging Microscopy (FLIM). The Soberman laboratory imaging suite includes 3 rooms dedicated to the study of protein interactions and cell biology using two systems for Fluorescence Lifetime Imaging Microscopy. These are time correlated single photon counting (TCSPC) systems. The first system, in a dedicated room, contains a Nikon TiE microscope interfaced with a 445nm laser and a Becker and Hickl FLIM system. The computer for this system is connected via a 1Gb Internet port to a remote server. The second system, in the adjoining room, is a TiE-perfect focus Nikon microscope with both epifluorescence and FLIM capabilities. This system contains two lasers for FLIM; a 448 nm and 515 nm. This system is also linked to the same server. Each laser contains a 60/40 beam splitter allowing swapping of lasers between FLIM systems without realignment. This configuration allows a highly flexible combination of a wide range of donor and acceptor fluorophores.
Time-lapse Microscopy. A time-lapse Nikon microscope with a heated motorized stage and a camera is located in the second room and interfaced to the server.
Data analysis. The third room contains three offline workstations linked to the server by 1Gb ports, allowing full data analysis at each station. The servers are linked to the Enterprise Research Infrastructure & Services (ERIS), which contains advanced data analysis programs for molecular imaging. A 4th room in the suite will be the future site of Super-Resolution microscopy.
N-STORM Super Resolution Microscopy. N-STORM (Stochastic Optical Reconstruction Microscopy). As described above, the 4th room in the suite will be the future site of Super-Resolution microscopy. In collaboration with Nikon Instruments and Micro Video Instruments, my group has access to a Nikon N-STORM (Stochastic Optical Reconstruction Microscopy). We will continue to have an N-STORM system in the imaging suite for a minimum of 8 weeks this year as last year).
Laser Capture Microdissection Microscope System. Now housed on the 6th floor will be present within the Soberman laboratory space due to formal interaction with the Neuroscience Microscopy Core.
• Additional Microscopy
Two Photon Microscopy for tissue-based FLIM. A BioRad Radiance Confocal Microscope configured for FLIM is available with unrestricted access for the Soberman lab on the MGH East campus in the adjoining building.
A TIRF Microscopy System-available through Neuroscience Core. A new Nikon A1R confocal microscope is available on the third (adjacent) floor. Dr. Soberman is the Chairman of the Advisory Committee for this facility.
• Protein Isolation and Analysis
The lab contains a large cell culture facility with three hoods, one of which is exclusively dedicated for the Soberman lab. The Soberman lab has four incubators, with one dedicated to SILAC (stable isotope labeling by amino acids in cell culture) for determination of proteins in macromolecular complexes by mass spectrometry and one dedicated to radiolabeling of proteins by 35S. A Biorad 2D protein electrophoresis and analysis system, and Typhoon scanner. This allows the use of DIGE labeling. Two HPLC systems are available for lipid analysis and protein analysis. The Soberman lab has an Applied Biosystems StepOnePlus for quantitative PCR.