We routinely prepare and analyse human (biopsies) and mouse bone tissues, both for static (on decalcified samples) and dynamic histomorphometry (on undecalcified samples). We also extend our histological expertise to some soft tissues (including frozen sample preparation and sectioning), upon request.
Bone samples follow a standardised processing protocol summarised below:
We use general as well as bone specific histological stains:
- Standard techniques such as haematoxylin and eosin, toludine and Alcian blue.
More specific stains such as Von Kossa and modified Glodner’s Trichrome (mineralised tissue), and tartrate-resistant acid phosphatase (TRAP) staining to identify osteoclast cells.
- We also use a range of fluorochromes to document bone deposition and remodelling on undecalcified samples.
|TRAP Staining of mouse bone||Von Kossa staining of mouse bone||Fluorescent quadruple labelling of mouse bone.|
|Goldner's trichrome of Human bone biopsy.||Fluorescent quadruple labelling of Human bone biopsy.|
Using two image analysis systems, we have the technology to perform static histomorphometric analysis to quantify parameters such as the number of osteoblasts and osteoclasts, bone surfaces and osteoid, as well as dynamic histomorphometry to measure bone formation rates using multiple fluorescent dye labels:
The Osteomeasure image analysis system has a manual tiling system allowing quantification in different fields of view, and is used for measuring e.g. TRAP stained slides or dual calcein labels on mouse bone sections.
The Bioquant Osteo system has an automatic microscope stage for accurate tiling across many slide fields, and is used specifically for human bone biopsy analysis.
Immunohistochemistry and Immunofluorescence:
Using paraffin tissue sections, a range of antibodies is available to identify and quantify changes in cells differentiation, distribution and activity, within the tissue; as well as monitor changes in the bone matrix (ex: bone development, bone healing). Detection can be chromogenic or with fluorescence.
|Osteoclacin - immunoflurorescence.||Ki-67 - Immunohistochemistry||Osterix - Immunofluorescence|