Most laboratories within LIMM have dedicated general purpose equipment rooms such as tissue culture facilities, dark rooms, radioactive and microbiology suites in addition to open laboratory workstations. These facilities are enhanced by state-of-the art facilities for in vitro and in vivo DNA, RNA and protein analysis.
The Institute runs a core funded DNA sequencing service based on an ABI 3130xi genetic analyser. This capillary sequencer allows accurate high-throughput sequencing of over 800bp in a single read, and microsatellite analysis for genetic linkage. High-throughput mutation analysis, genotyping and SNP discovery is also possible using an Idaho Technologies Inc. LightScanner.
Two robotic workstations, including a Biomek 2000 Liquid Handling Robot for automated pipetting, diluting and dispensing operations, are available for high throughput PCR.
Multiple Real Time (ABI 7700 and ABI 7500) PCR machines enable, high-throughput, accurate measurement of transcript expression levels from cells and tissues. These include a micro-fluidic card ABI7900, 384-well plate system, compatible with fully automated robotic loading. This facility is complemented by RNA extraction platforms (ABI6100 DNA and RNA preparation station, and nanodrop analyser) that can be used in combination with on-site laser micro-dissection microscope facilities (Pixell Laser Capture with microdissection and microablation capabilities).
LIMM possesses excellent facilities for immunocytochemical and immunohistochemical microscopy imaging with extensive slide scanning-imaging and storage capacity. These include the newly developed Nikon Eclipse confocal microscopy system allowing the 3D visualisation of cell or tissue structure and the analysis of protein expression patterns and intracellular localisation. It incorporates four laser sources, and is built around an inverted microscope with an environmentally-controlled stage, allowing the imaging of living samples. The system is also equipped for Total Internal Reflection Fluorescence (TIRF) microscopy, allowing very high resolution imaging of fluorescence closely associated with the cell base.
Two systems are available for time-lapse imaging of living cells by either fluorescence or bright field microscopy. One is equipped with an Eppendorf micro-injection system while the other has a motorised stage allowing automated high-throughput multisite imaging. A variety of image analysis software, including deconvolution options, is available for these systems.
Numerous other systems are located throughout LIMM for routine fluorescence microscopy and image capture. These are complemented by an additional Zeiss Palm microbeam Laser Capture Microdissection microscope and a Vysis Quips Xl FISH Analysis system.
A state of the art MoFlo Cell Sorter (Dako Cytomation) for high speed cell sorting, and a FACS-Vantage cell sorter is provided, as is a 7 colour flow cytometer with red, blue and UV lasers (Beckton Dikson LSR II cytometer), and several other flow cytometers for 3 and 4 colour analyses (FACS-Calibre, FACS-Scan, Guava Easycyte). This is a core facility, managed by dedicated technical support
Two Bio-Rad Pharos FX Plus Molecular Imager-S phosphorimager systems are available for the detection and expression analysis of DNA, RNA, or protein samples in gels, blots, or microplates. Each laboratory is also equipped with Bio-Rad Gel Doc XR imaging systems for routine image capture of dye stained agarose and polyacrylamide DNA and protein gels. A Chemi-Doc XRS system enables image capture and quantitative analysis of chemiluminescent images, often useful for Western blot analyses of protein expression in cell extracts. Alternatively, a Licor 9201 Odyssey Infrared Imaging System may be used for quantitative Western or Northern blot analyses.
The presence of several flow-based ELISA systems (Luminex Technology) permits quantification of soluble molecules such as cytokines, phosphorylation signalling cascades, and DNA-protein complexes.
The Institute uses the central University of Leeds email and data storage servers as well as running its own SAN. The Institute SAN consists of a number of Apple Xserve servers, Promise and Apple RAID arrays all backed up by an online Sun Virtual Tape Library (VTL) system and offline to magnetic media. The Xserve servers also provide other services to the Institute such as web site hosting, secure enterprise video conferencing and Filemaker Pro server hosting. The Institute also has a number of high performance computing systems run by a team of bio-informaticians. Between the numerous sections of the Institute there are over 700 PCs and Apple Macintosh computers all of which is supported by a team of dedicated IT technicians.
The Institute jointly operates an Illumina GAIIx clonal sequencer and associated equipment and reagents enabling genome-scale or targetted sequencing projects to be executed in a highly cost effective manner.