At the University of Portsmouth we have been able to establish an excellent suite of laboratories with tissue culture laboratories and a host of state-of-the-art equipment including those to enable various assays including histology, flow cytometry, cellular oxygen utilisation measurement, ECIS, TEER/Flocell/Cellix, Auto-MACs cell separation, cell viability, apoptosis & proliferation and RT-PCR, metabolic, gene and proteomic analysis which we have integrated into dedicated histology, cellular, molecular laboratories respectively. We have also established a sophisticated microscope imaging suite which includes atomic force, laser capture micro-dissection, confocal/live cell, total internal reflection (TIRF), Ariole quantitative image analysis and micro-fluidics/live cell microscopy as well as having full access to scanning and transmission electron microscopy and confocal microscopy. We have also developed a human biopsy-derived brain tumour cell culture repository holding well characterised (gene and protein levels), DNA fingerprinted, mycoplasma tested cells with an on line registry which we share with our collaborating laboratories without the UK.
Housing the Charlie’s Challenge room with epifluorescence/phase/brightfield Zeiss microscope, the Luke Frost room for establishing biopsies in culture and the Diana Ford room for generic tissue culture (each is fully equipped and with hypoxic and normoxic incubators). The Diana Ford room also houses Flocell, EVOHM/TEER, Electrical Cell Impedance Sensing and CellZscope physiological monitoring of TEER, cancer metastasis and nanoparticle delivery through our 3D BBB models. A BD FACScalibur four-colour Flow cytometer, a Chemometec Nucleocounter NC-3000 fluorescence imaging cell analyser for cell cycle, viability and apoptosis, a series of 4 Oxytherm electrodes (Clarke type) for oxygen uptake measurements on mitochondrial and cellular respiration, a BMG Labtech POLARstar OMEGA plate reader with atmospheric control unit for live-cell analysis incorporating normoxic or hypoxic conditions, a microplate reader with fluorescence intensity (including FRET), polarization, luminescence (including BRET), UV/Vis absorbance, and time-resolved fluorescence (including TR-FRET), and AlphaScreen®/AlphaLISA®, a BMG Labtech POLARstar OPTIMA Plate reader using fluorescence, luminescence, and absorbance detection modes, a Countess II Fl automated Cell Counter with fluorescence, an EVOS FL Auto Live-Cell inverted digital imaging system providing three-colour fluorescence images or videos in a controlled on-stage incubator and a three fluorescent channel Floid Cell Imaging Station are also located here.
We opened The Alison Phelan Memorial Laboratory in October 2012. The lab now houses molecular equipment, e.g. Western blotting (Bio-Rad MinoPROTEAN, Pierce PowerBlotter), PCR (Bio-rad MyCycler) etc, a 2100 Agilent Bioanalyser providing sizing, quantitation and quality control of DNA, RNA, proteins and cells (used in DNA fingerprinting and RNA integrity), a Licor Odyssey CLx Infrared Imaging System (can quantitate two targets on same blot simultaneously) and a computer with SmartType Chromosome/karyotyping software.
With fume cupboards, Leica EG1150C FFPS processing/embedding stations, 3 rotary paraffin and 1 resin section Leica microtomes, a Leica CM3050C cryostat and tissue slicer.
Microscope Imaging Suite
Housing a Bruker Multimode and Nanoscope Atomic Force Microscope, a Leica Ariole brightfield, fluorescence and FISH slide microscope system with image analysis software, a Zeiss Total Internal Fluorescence Microscopy (Evanescence laser wave technology)/SIRF microscope, a Zeiss Inverted live-cell imaging microscope with environmental chamber for hypoxic and normoxic culture and Cellix microfluidics system, an AxioImager Z1 Fluorescence Inverted microscope with micro-injection, x2 Zeiss Laser confocal microscopes (LSM 510, LSM 710), an Arcturis/Veritas Nikon laser capture microdissection scope, an EVOS live cell systemand a high performance computer for mitochondrial DNA/3D modelling. Dr James Smith (a HEFCE funded SRF) manages the facility.
Electron Microscopy Suite
In addition, we have access to an electron microscopy suite which houses ultra microtomy and scanning and transmission electron microscopes, and associated biological equipment.