Histology

Automated Slide Handling

The Histology unit provides a range of services available to researchers within the Institute. Unstained high quality paraffin / frozen sections are produced for immunohistochemical analysis, in situ hybridisation and laser capture microdissection. Alternatively the sections may be stained by H+E for morphological examination or by more specific special stains. We also provide a comprehensive immunohistochemistry staining and troubleshooting service. Methods are also available for the preparation and staining of cells (cultures, cytospins, smears and pellets). Facilities are available for light microscopy, heat induced epitope retrieval, counterstaining and coverslipping, as well as advice and protocols on any area of histology.

The fully automated Mirax Scan system for high throughput scanning of histological preps, resulting in highly resolved digital images of a whole slides is available in advanced imaging unit

Fixation, processing and microtomy

No one fixative is ideal, and as result, the unit provides a selection of the most commonly used fixatives, together with advice for correct choice. Specimens are processed on the automated Shandon Excelsior tissue processor using various schedules depending upon the size and nature of the tissue. Paraffin sections may be stained for morphological examination using the Leica autostainer XL, or unstained for protein extraction, immunohistochemical analysis or in situ hybridisation.

Cryotomy

Processing methods can have a deleterious effect on tissue constituents, and as a result, in some circumstances frozen sections may be the preferred method. Advice on all aspects of frozen tissue preparation and post fixation is available. The preparation of fixed cryoprotected samples is also routinely performed.

Special Stains

Special stains allow for the demonstration of a variety of tissue constituents. Common requests include lipid, carbohydrate and cytoplasmic granule staining, together with various single and trichrome techniques for connective tissues and extracellular substances. Methods for specialised haematoxylin, nucleic acid and bone preparations are also available.

Immunohistochemistry

Immunohistochemistry is a technique employed by many groups within the Institute. The unit houses the i6000 automated IHC platform and several epitope retrieval stations available for use.

Advice and support in all areas such as choice of fixation, epitope retrieval, method design / deployment and visualization is available.
Optimisation of novel antibodies, multiple chromogenic and fluorescent labelling studies, troubleshooting and interpretation of results are commonly undertaken.

 

dual labelling image

Dual labelling using Ki67 (brown) & CD31/34

 

i6000 platform

i6000 platform

 

In Situ Hybridisation

This is an established, available technique. Advice on section preparation, fixation and pre-treatments, plus manipulation of the probe hybridization, stringency washing and probe detection to overcome the unique problems associated with in situ hybridisation is available. Optimisation of novel probes and troubleshooting are often undertaken. The Hybaid Omnislide system is available.

melastatic melanoma
Metastatic melanoma mRNA in situ hybridisation

melastatic melanoma
Metastatic melanomaDa H&E

 

Laser Capture Microdissection

An available technique which allows individual cells from specific microscopic regions of tissue sections to be isolated for molecular analysis. Although paraffin sections afford better morphological preservation, frozen sections give excellent recovery of both DNA and RNA. Cell identification can be either by morphological analysis or based on immunophenotype.
The facility houses the Leica LMD6000 system which with an integrated scanning stage and automated cell recognition facility can provide high sample throughput, whilst reducing sample preparation time.

LMD6000
LMD6000

 

Endothelial cells
Endothelial cells highlighted on an ovarian tumour.
Shown before and after dissection.

 

Karyotyping

A routinely provided service which after manipulations to introduce DNA into ES cells, the karyotype of the cells is checked to ensure they have a normal karyotype and hence are able to contribute to the germline of the chimeric mouse.

Tissue Microarrays

Tissue microarrays allow for high throughput analysis and standardisation of methodologies, whilst also preserving rare samples. Both immunohistochemical and molecular detection techniques can be performed on TMA slides.
Tumour-specific, control and custom arrays are routinely constructed. Up to 400 cores can be transplanted to the recipient block.
The unit houses the Beecher MTA1 manual arrayer and due to increased demand the automated Beecher ATA27 will soon be available.

ATA27
ATA27

 

Donor Cores
Recipient block containing donor cores

 

Manchester Cancer Research Centre Biobank

The Histology Facility also houses the Manchester Cancer Ressearch Centre Biobank, an initiative established to facilitate an organised, systematic and comprehensive collection of high quality samples from cancer patients across five different Trusts in the Manchester area.

Developments in genomics and proteomics have highlighted the need for large numbers of tissue and blood samples to be made available to facilitate translational research within the Manchester Cancer Research Centre.