Histopathology and in situ hybridisation core facility

The Histopathology/in situ hybridisation (ISH) core facility at the Cambridge Research Institute offers a variety of histological techniques, immunohistochemistry, in situ hybridisation, laser capture microdissection as well as automatic slide digitisation and analysis to CRI scientists.

Histology

The facility processes, embeds and sections human and animal tissues or cell lines into frozen or paraffin formats and stains these with the standard haematoxylin and eosin (H&E) or special stains, as needed by the researcher to complement their work. During the past year, the facility has worked up further special stains, the Herovici stain for new deposition of collagen and the Leder stain for mast cells (Figure 1), which can be added to the list of special stains being run by the facility.

Mast cells stained with the Leder stain within the interstitium of a papilloma (Histopathology core facility report 2010; figure 1)
Figure 1
Mast cells stained with the Leder stain within the interstitium of a papilloma.

Immunohistochemistry (IHC)

A third Bondmax was purchased last year for antibody work-up and has contributed to an increase in the number of available antibodies for routine staining (170), and to a 51% increase in the number of slides stained by the facility this year to 20,000. We are currently undertaking a project in collaboration with the Caldas laboratory to assist in the profiling of 150 antibodies across 3,000 breast cancers in tissue microarrays. Dual fluorescence has also been worked up on the system (Figure 2).

Dual immunofluorescence of Cytokeratin18 and Androgen Receptor in normal human prostate (Histopathology core facility report 2010; figure 2)
Figure 2
Dual immunofluorescence of Cytokeratin18 and Androgen Receptor in normal human prostate.

In situ hybridisation (ISH)

Our gold standard ISH protocol utilises S35-labelled riboprobes and we offer fluorescence ISH (FISH) for the Y chromosome and HER-2 CISH as part of the routine service. In collaboration with the Caldas laboratory, we have worked up a micro-RNA ISH protocol that is working well for high-medium abundance targets. Our aim in the next year is to apply this in a reconditioned Bondmax system that has been purchased for this purpose.

Digitisation and analysis

The Zeiss Mirax system, purchased last year, has increased the facility's capacity for digitising brightfield and fluorescence slides, raising our scanning output by 39% to 30,000 slides. We have continued to assist users in maximising the use of the analyses that can be carried out (Bolton et al., Cancer Epidemiol Biomarkers Prev 2010; 19: 992) and have been involved in a major collaboration with the Breast Cancer Association Consortium (Sherman et al., Cancer Epidemiol Biomarkers Prev 2010; 19: 966). The numbers of slides being captured for image analysis has steadily increased and we hope to grow this further.


Facility manager
Will Howat is the head of the Histopathology and in situ hybridisation core facility.