MCB faculty research is supported by a variety of centralized instrumentation/service facilities:
- Microscopy and Imaging Center
- High Field NMR Facility
- Mass Spectrometry and Molecular Weight Determination
- Macromolecular X-ray Diffraction Facility
- UMass Zebrafish Facility
- Growth Chambers in the Greenhouse Facility
- Flow Cytometry Facility
- Center for Excellence in Apoptosis Research
The university's Central Microscopy Facility offers state-of-the-art equipment and expertise, with emphasis on biological applications. It is organized so that students and faculty can obtain advice on their projects and training on the instruments they will need to use. Once training is complete, the facility is available round the clock.
For light microscopy applications, a Nikon Diaphot 200 provides DIC and fluorescence optics, as well as automatic photomicrography and image digitization using a Diagnostic Imaging Spot-RT camera. This system features automated fluorescence excitation and emission filters and Z-focus for widefield image stacks in up to three channels. A Zeiss 510-META confocal microscope is also available for critical work in which depth resolution or 3D reconstruction is desired. The confocal microscope is a state-of-the-art system allowing simultaneous collection of three fluorescence channels and DIC as well as 32 channel spectral imaging, FRET, FRAP, and other methods. Transmission electron microscopy (TEM) is available through a computer controlled FEI Tecnai T12 with digital camera and CompuStage, or JEOL 100s instruments. For scanning electron microscopy (SEM), the facility operates a JEOL JSM-5400 with digital image acquisition.
There is also a complete range of equipment for the preparation of biological materials for light and electron microscopy. Low-temperature preservation is especially well represented, with Balzers freeze-fracture and liquid propane plunge freezing and low-temperature substitution. A cryo-holder is available on the FEI T12 TEM. Computer image processing facilities available in the Microscopy Facility include a dedicated workstation with software packages (Media Cybernetics "AutoDeblur" and "Autovisualize" products) for deconvolution of widefield, confocal, and DIC images and 3D rendering, measurement, and analysis (AutoDeblur and Autovisualize products) of volume data.
The High Field NMR Facility is jointly operated by the departments of Biochemistry and Molecular Biology, Chemistry, and Polymer Science and Engineering, under the direction of a Ph.D.-level scientist. There are two high field spectrometers available for routine analysis (DPX300, Avance400 spectrometers manufactured by Bruker).
Three additional spectrometers are available for advanced applications. The Avance600 spectrometer has a cryoprobe and is fully equipped for triple resonance and gradient experiments, while the DSX300 and Infinity300 are wide bore solid-state spectrometers equipped for triple resonance experiments as well as high-speed magic angle spinning, static solids, and wideline experiments.
The advanced spectrometers are capable of the latest techniques for determining the structure of macromolecules, including multinuclear, multidimensional NMR analysis of proteins and site-directed studies of insoluble, larger systems such as membrane proteins.The spectrometers are Ethernet-linked to a number of Silicon Graphics and Windows computers for data analysis, computation, and manipulation of 3D structures.
The Mass Spectrometry Facility is one of the best equipped in the area, with the following instrumentation:
- Esquire-LC electrospray ion-trap (Bruker Daltonics, Inc.) equipped with HP1100 HPLC system. This system is designed for LC-MS applications, but can also easily be used for analysis of peptides, proteins, and other biopolymers. With capabilities of MSn, it is also a valuable structural tool.
- Reflex III MALDI-TOF (Bruker Daltonics). The "unlimited" mass range of a time of flight mass spectrometer enables the molecular weight and polydispersity of polymers to be analyzed.
- JMS700 MStation (JEOL) high resolution two-sector mass spectrometer. This instrument is equipped for EI, CI, FAB, ESI, and FD ionization methods.
- MS Engine (Hewlett-Packard) GC-MS system. This system can be used for separation of complex mixtures by gas chromatography and identification by mass spectrometry.
Complementing the existing structural biology facilities, the newly opened X-ray diffraction facility allows researchers to solve macromolecular structures by X-ray crystallography. The facility consists of a RigakuMSC RU-H3R X-ray generator and RAXIS 4++ X-ray detector equipped with Osmic Confocal Max-Flux optics, an inverse phi goniostat, and an Oxford cryojet. With the support of two crystallographic laboratories in the biochemistry department (Dr. Theis, Dr. Garman) and one in the chemistry department (Dr. Hardy), the facility is able to provide training, technical expertise, instrumentation and computational infrastructure for obtaining well-diffracting crystals, mounting crystals, measuring high-quality diffraction data, processing data, solving structures by molecular replacement or MIR/MAD techniques. For example, users have access to a dynamic light scattering instrument housed in Dr. Theis's laboratory for screening samples for crystallizability.
Contact: Judy Bennett (firstname.lastname@example.org)
The Biology Department has three research groups that study embryonic development using the zebrafish as a model system. Each group maintains a separate zebrafish room. Up to 30,000 fish are swimming around at any given time.
Contact: Elsbeth Walker (email@example.com)
The growth chambers, supplied by the Conviron Company, are located in a room of over 1000 sq ft. that is contiguous with ranges #1 to #4. The equipment includes: Four PGW-36 large reach-in units with 3.3 m2x 200 cm of growing space. Lighting delivers 960 umoles/m2/sec, and temperatures may be controlled between 4 and 45C with a variation of 0.5C.
Four E7/2 dual compartment units, each of which has 0.76 m2 x 64 cm of growing space. Lighting delivers up to 33 umoles/m2/sec. Temperature ranges are similar to the reach-in chambers. (Four other E7/2 units were also purchased at the same time, but these are located in the Lederle Graduate Research Tower, home of the Department of Biochemistry and Molecular Biology.)
One 125L incubator with five shelves, lighting and humidity controls, and horizontal airflow patterns. All of the growth chambers feature an adjustable, counterbalanced light canopy, a sampling tube that allows control at plant canopy height, and a design that permits plants to be placed directly on the floor.
Environmental control: Temperature, lighting, and humidity are controlled by the Conviron CMP4030 Operation System, a dedicated computer located in the headhouse. The computer allows multi-day programming and all functions are performed by touching the back-lit, high resolution, VGA liquid crystal display. The program also allows graphing of set point and actual performance, so there is no need for a chart recorder.
The Flow Cytometry Facility includes a two laser LSR II flow cytometer and a FACSVantage SE cell sorter.
The FACSVantage SE is configured with 488nm and 633nm air cooled lasers. This FACSVantage can perform eight parameter analysis and sorting. The MacroSort option on the instrument allows large particle analysis and sorting.
The LSR II is configured with 488 nm and 633 nm fixed alignment lasers. The primary blue 488 nm laser detects five fluorochromes while the red 633 nm laser detects three fluorochromes.
CEAR is based in the Pioneer Valley Life Sciences Institute's state-of-the-art biomedical research space in Springfield, Massachusetts. General laboratory space is designed to maximize scientific interaction among diverse researchers. It is fully equipped and staffed for animal care, histology, cell culture, and molecular biology. Two surgical suites are available for work with animal models to facilitate training, research, and medical instrument development. Shared equipment includes:
- FACScanConfocal laser microscope
- Whole animal in vivo imaging system
- Real-time PCR Robotics devices for conventional histology and immunohistochemistry.