With the aims of sharing information about the technical aspects of immunohistochemistry
(IHC) and making it possible to make a suitable choice of antibody for histopathological
examination, this technical report describes the results of a questionnaire administered
during the period of 2014 to 2015 to members of the Conference on Experimental Animal
Histopathology. It also describes the immunological properties of primary antibodies
(clone, supplier, catalog number, species reactivity, etc.) and the IHC staining
conditions (fixing solution, fixing time, embedding, antigen retrieval method, antibody
dilution, incubation time, incubation temperature, positive control tissue, secondary
antibody information, etc.) for a total number of 733 primary antibodies (425 kinds of
primary antibody).
A subcutaneous mass in the right femoral region of a female F344 Slc/N rat was examined
histopathologically. At 83 weeks of age, the animal showed symptoms of severe anemia and
nasal bleeding. Necropsy revealed that the mass had invaded the skeletal muscles but did
not affect the bones. Multicentric nodules were also observed in the lung. Histopathology
revealed a sheet-like growth pattern of polygonal tumor cells with round or comma-shaped
nuclei and pale eosinophilic cytoplasm. Osteoid tissue was observed in not only the
original lesion but also the metastatic foci in the lung. Each tumor cell was surrounded
by argentophil fibers and few collagen fibers. Immunohistochemically, the tumor cells were
positive for proliferating cell nuclear antigen (PCNA), vimentin, osterix and osteocalcin,
but negative for keratin, S-100, von Willebrand factor, CD-31, CD-34, desmin, α-smooth
muscle actin, lysozyme, α1-antitrypsin and rat malignant fibrous histiocytoma (MFH)
antigen. CD-68-positive cells were considered to be infiltrated macrophages because they
were negative for PCNA. On the basis of these findings, we diagnosed the present case as
extraskeletal osteosarcoma.
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