Intermediate Filament Staining in the Cytologic and Histologic Diagnosis of Canine Skin and Soft Tissue Tumors

Abstract

Abstract. Formalin-fixed histologic and acetone-fixed cytologic preparations from 87 surgically removed subcutaneous and soft tissue canine tumors were examined for immunoreactivity to cytokeratin, desmin, and vimentin. The avidin-biotin complex (ABC) technique demonstrated immunoreactivity in both preparations, but the intensity and specificity o~t~ on t he mimary antibody. __ Polyclonal antibodies to cytokeratin were more consislentinimmunoreactivity than were polyclonal desmin or vimentin antibodies. ThFmTnoclonal antibody proved more s a t~~~t o r y f o r~d e~n s t r~t i~~v i m~~~~~~a n the polyclonakmtibody. Greater dilutions of primary antibodies may be used on cytologic preparations than on histologic sections. Evaluation of cytologic preparations may be inconclusive due to background staining, scant cellularity, or poor cytoplasmic preservation.Intermediate filaments have been used as cell typespecific markers for histologic and cytologic evaluation of normal and pathological t i~s u e s .~J~J~,~~ Antibodies that discriminate between different proteins of intermediate filaments can be used to identify tumor cell type and o~i g i n .~J~,~~ Cytokeratin, vimentin, and desmin are present in epithelial, mesenchymal, and muscle cells, respe~tively.~~,~',~~,~' The purposes of this study were to apply avidin-biotin immunoperoxidase staining techniques to demonstrate these proteins of intermediate filaments in tissues and to evaluate the accuracy of these antibody techniques in the diagnosis of routine cytologic preparations. Materials and MethodsEighty-seven neoplastic masses were selected for immunoperoxidase staining in a prospective study from May 1986 to July 1987. Cytologic aspirates or imprints were obtained from surgical tumor biopsy specimens from canine patients. Cytologic preparations were air-dried, fixed in cold acetone for 10 minutes, and stored at -40 C. Surgical biopsy specimens were fixed in 10% neutral-buffered formalin. One cytologic and one histologic preparation were stained with Wright's stain and hematoxylin and eosin (HE), respectively. The remaining unstained slides were used for immunoperoxidase staining by the avidin-biotin-peroxidase complex (ABC) method (Vectastain ABC Kit).15-17 For histologic specimens, paraffin-embedded tissues were sectioned at 5 pm, deparaffinized, rehydrated, and incubated with 0.1% trypsin and 0.1% CaC1, pH 7.8 for 15 minutes at 37 C.7J8 Sections were rinsed in phosphate-buffered saline (PBS) for 10 minutes and placed in 0.3% hydrogen peroxide in methanol for 20 minutes to remove endogenous peroxidase activity. After a 20-minute rinse in PBS, sections were covered with 1% normal goat serum for cytokeratin and desmin staining and 1% rabbit serum for vimentin staining. This was followed by incubation of one slide each with rabbit

Haemangiopericytoma: histological spectrum, immunohistochemical characterization and prognosis

“…In addition to smooth muscle actin, six CHPs co‐expressed desmin: four with perivascular whorled pattern and two with storiform pattern. Desmin has been detected previously in neoplastic cells of CHPs: this marker was co‐expressed in 1 of 23 muscle actin‐positive CHPs 8 in one of two CHPs studied by Andreasen et al 26 . and in the unique case described by Thoolen et al 27 .…”

Technical Considerations for Developing Enzyme Immunohistochemical Staining Procedures on Formalin-Fixed Paraffin-Embedded Tissues for Diagnostic Pathology

“…The ABC immunostain has been used for the detection of most tumor markers and infectious disease agents in formalin-fixed tissues. [1][2][3][4]822,23,28,29,37 Figures 4a and 4b demonstrate ABC immunohistochemical detection of rabies and bovine viral diarrhea virus, respectively, in 27 Processed tissue in paraffin blocks may be stored indefinitely at room temperature without further loss of antigenicity. We have immunohistochemically detected bovine respiratory syncytial virus antigens in a section cut from a paraffin block stored for over 30 years.…”

The Spectrum of Canine Cutaneous Perivascular Wall Tumors: Morphologic, Phenotypic and Clinical Characterization

“…Based on immunohistochemical staining, a phenotypic continuum from HEP to adventitial tumors was evidenced in 16 dogs (dog Nos. [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16]. Contractile protein expression (Table 3) was minimal in the HEP; the amount increased in the group of myopericytomas, was maximal in the myomas, and was progressively reduced among the adventitial neoplasms recapitulating the phenotypic expression of these proteins in the normal vascular wall cell types.…”