100 x magnification.
The slide on the left is a 5 micron H&E stained section from the gross slabbed block of a femoral head / proximal femur / tumor composite specimen. The resected specimen was gross cut on a band saw into 5 mm thick slabs prior to formic acid decalcification and paraffing processing. The 120 mm long block was sectioned via a Lipshaw 80A manual sledge microtome according to established protocol . (J Histotechnology 20(3):267-277, 1997)
This represents a 5 micron thick section of a mature bone - reparative bone interface in a fracture model. The specimen was decalcified in 5% formic acid prior to paraffin processing. This section is stained with a Gill’s II Hematoxylin cellular stain and an Eosin Y / Phloxine counterstain (H&E). This staining procedure is the foundation for the majority of histologic studies performed in The Center.
This consecutive serial section of the field in the above H&E is stained with a Safranin O – Fast Green (SOFG) procedure. The Safranin O stain stains proteoglycan and is useful in enhancing any cartilage which may be present and not easily identifiable in the H&E stain.
(Trichrome Mature Bone – New Bone)
The Masson Trichrome performed on the consecutive serial section of the
above SOFG field is useful in enhancing connective tissue as well as
improved photographic contrast when compared to the H&E stain.
(Gram Stain)
Studies of Osteomyelitis and infectious Arthritis are a significant part
of The Center’s focus. This decalcified, paraffin processed section of
infected bone was stained with our modified Gram Stain illustrating the
presence of a known strain of Gram Positive Bacteria (blue – black)
embedded deep in the bone (red) . (J.
Histotech., 15:303-306, 1992.)
(von kossa)
In addition to the above decalcified paraffin histology, The Center is
equipped to do Glycol Methacrylate (GMA) histologic processing enabling
the investigator to study undecalcified sections. Depicted here is a two
micron section of a healing bone model sample, processed undecalcified in
GMA and stained via a modified Von Kossa method. The black lines denote
mineralization within the field (J. Histotech., 16:129-137, 1993).
In addition to the above GMA processing, the Center for Orthopaedic Research is also expert in undecalcified methyl methacrylate (MMA) processing and histomorphometry. Depicted here is a four micron section of fetal bone, processed undecalcified in MMA and stained with Masson's trichrome.
