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WORLD CLASS BASIC, TRANSLATIONAL AND CLINICAL RESEARCH

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Instrumentation

High Content Screening

3D organoid acquired on the Opera Phenix using plastic bottomed 96-well plate with the 20x water objective in confocal mode. The organoid is stained with DAPI in blue and an membrane marker in green.

3D nuclear detection using PerkinElmer Harmony is demonstrated.

Data from Christopher Below and Claus Jorgensen (Systems Oncology).

Allowing the imaging and analysis of the phenotype of multiple cell populations so to study cellular responses, for applications such as drug discovery, RNAi, and 3D culture (spheroids and organoids).

  • PerkinElmer Opera Phenix, a spinning disk based confocal screening system with a dedicated sCMOS camera per detection channel (4). Magnification from 1.25-x60 (water). Automation allows loading of multi-well plates at room temperature or from an incubator.
  • PerkinElmer Operetta CLS, a fluorescence wide-field based illumination system for screening system multi-well plates

Super Resolution Microscopy

Fluorescent Histone H3 in nucleus of a HeLa cell visualised under confocal microscopy (left) and super resolution microscopy using gSTED (right).

Data from Steve Bagley.

The aim of this equipment is to achieve a resolution and clarity suitable for the task of visualising cancer cells at the molecular level.

  • Leica gated Stimulated Emission Depletion Microscopy (gSTED) with additional confocal and multi-photon illumination. Under gSTED illumination and image deconvolution a resolution of 30nm can be attained.
  • Zeiss LSM880 Airyscan, temperature and gaseous control permit the researcher to image live cells under low laser light illumination to a resolution of 80nm

Confocal and Widefield Microscopy

Confocal microscopy image of a single cell stained for Factin using Phalloidin.

Data from Steve Bagley.

Imaging volumes of data via laser illumination so to examine the 3D nature and dynamics of cells and tissues.

  • Zeiss Airyscan (LSM880, invert stand), laser scanning confocal and fast super resolution. Fitted with an additional 355nm laser for induction of DNA damage.
  • Leica (SP8 MP CSU, upright stand) confocal, two photon confocal microscope with second harmonic imaging for the visualisation of tissue and 3D cell culture
  • Leica (TCS SP8 inverted stand) confocal and two photon microscope with second harmonic imaging for the visualisation of tissue and 3D cell culture
  • Leica Macro-Confocal (TCS LSI) when imaging a large field of view is required such as primary tissue for translational studies
  • API Deltavision Core
  • Low light imaging system with an Andor iXon 888 camera based around Metamorph

Histology Scanning (Virtual Microscopy)

Benign Prostatic Hyperplasia - DAPI panCK, RunX1, RunX2.

Using RNAscope, whole slide imaging and HALO detection of localized mRNA in intact cells. Using these technologies a study of the molecular mechanisms of the cell is possible.

Data by Renaud Mevel and Georges Lacaud (Stem Cell Biology).

Automated digitisation of whole histology slides for the visualisation of multiplexed tissue (bright-field and fluorescence) at x400 total magnification. Image analysis and statistical analysis is provided offline via the Definiens (tissue studio and image miner) and the PekinElmer Inform (Vectra) software.

  • Leica SCN400, bright-field illumination, able to batch up to 250 slides
  • PerkinElmer Vectra 3 automated multi-spectral imaging system, bright-field and fluorescence, able to batch process up to 150 slides
  • Leica Versa, bright-field and fluorescence illumination, with a range of objective lenses from x4-x63, allows automated oil immersion for high resolution imaging. Able to batch process up to 150 slides

Radiation Sources

Xstrahl CIX3 system for x-ray irradiation of cells in a variety of vessels (misc. tubes, well plates, petri). The radiation quality of the output has a half-values layer of approximately 2.3mm Cu.

Miscellaneous Equipment

  • The SeaHorse XFe Analyser, fast real time measuring of mitochondrial respiration and glycolysis in a microplate
  • Makerbot Replicator. 3D desktop printer for building customised equipment and pre-production manufacture

Software

A variety of image processing and analysis packages are available along with IT infrastructure (1 Linux and 1 windows multi-processor system, eight workstations, 200TB of temporary storage via the Scientific Computing department).

  • Histological Image processing and Analysis: HALO: Definiens Tissue Studio: Definiens Image Miner, PerkinElmer Inform/Phenochart, Bitplane Voloom
  • High Content Screening: PerkinElmer Harmony , Columbus, CellProfiler
  • Microscopy: Bitplane Imaris, Huygens, Matlab, ImageJ/Fiji