Experimental Histopathology

Cell and tissue phenotyping is fundamental to our understanding of molecular, cell and tissue mechanisms in developmental biology, cancer and infectious disease. The Experimental Histopathology Laboratory provides a comprehensive, interactive centre for experimental tissues and human bio resources/clinical trials, with the ultimate aim of relating pathology of rodent and other animal models of disease to that in humans.

We provide expert histopathological phenotypic analysis; identifying strain versus mutation-specific changes, specialist interpretation of tissue structure including advice on embryonic development, human comparative pathology and advice on experimental designs for future studies, reducing the burden on researchers and consistent with the guiding principles of refining, reducing and replacing the use of animals in scientific research.

Technical procedures performed include:

  • Optimal handling, processing and sectioning of a wide range fresh, fixed and frozen tissues, with specialist dissection of adult rodents and embryos (Nikon SMZ 1500 for dissecting microscopy)
  • Mouse developmental analysis including embryonic lethal phenotypes
  • Human and rodent histopathology
  • Slide evaluation using MicroPix Quad image capture teaching/discussion platform with on-site pathologist
  • Whole-mount or tissue section histochemistry and immunohistochemistry (IHC) and non-isotopic in situ hybridisation, with the advantage of a Ventana Discovery XT IHC/ISH automated system for high throughput. Novel antibody optimisation for mouse/human IHC (access to human/rodent control tissue and a database of >350 optimised antibodies)
  • RNAscopeTM ISH technology, a highly specific technique for demonstrating even extremely low abundance mRNA for any gene, from any species, in routinely fixed (FFPE) tissue.
  • 3D reconstruction - Bitplane Imaris 8 software enables for 3D volume rendered reconstruction of precisely captured images from serially stained tissue sections, for a 3-D view of cellular relationships and tissue architecture
  • High resolution photomicroscopy and morphometric analysis for bright field and fluorescence tissue sections using a Nikon 90i microscope with NIS Elements software platform for semi-automated object classification/measurement.
  • Laser Capture Microdissection (Leica LMD 7000) - allows for contact and contamination-free isolation of specific regions of interest down to single cells or cell components to be isolated, via bright field or fluorescence illumination, with the Automated Vision Control (AVC +) software for automated cell recognition.
  • DNA/RNA extraction from FFPE
  • High quality scanning microscopy using the Ariol SL50 and the new Zeiss Axio Scope.Z1 scanner
  • Morphometric analysis using Genetix morphometric software with:
    • IHC module for quantitating membrane, nuclear and cytoplasmic expression
    • Fluorescent module for FISH and immunofluorescent capture
    • Tissue Microarray module
    • General Morphometric Image Analysis module (ploidy, angiogenesis, area/volume etc.)

The EHP promotes a close interaction with the Biological Resources facility, research groups and other STPs. Scientists are encouraged to work in our laboratory and collaborate on experimental design, to conserve time and effort and advise where other STPs may optimise the strategy. Teaching is an essential activity and individual training in animal/embryo dissection, photomicroscopy, tissue handling, IHC/ISH and other laboratory techniques is provided.

Emma Nye (Lead) 

emma.nye@crick.ac.uk
+44 (0) 20 7269 3621