Quantification of tumour vascularity in squamous cell carcinoma of the tongue using CARD amplification, a systematic sampling technique, and true colour image analysis

• Published in: Analytical cellular pathology 2001, Vol.22 (4), p.183-192
• Main Authors: Hannen, E J, van der Laak, J A, Kerstens, H M, Cuijpers, V M, Hanselaar, A G, Manni, J J, de Wilde, P C
• Format: Article
• Language: English
• Subjects: Antigens, CD34 - biosynthesis; Biotinylation; Carcinoma, Squamous Cell - blood supply; Genetic Techniques; Humans; Image Processing, Computer-Assisted; Immunoenzyme Techniques; Immunohistochemistry; Lymphatic Metastasis; Neoplasm Metastasis; Neovascularization, Pathologic - pathology; Observer Variation; Other; Reproducibility of Results; Tongue Neoplasms - blood supply; Tyramine - metabolism
• ISSN: 0921-8912; 2210-7177; 2210-7185; 1878-3651
• DOI: 10.1155/2001/780576

The aims of this study of head and neck tissue samples were to develop an immunohistochemical protocol based on the catalysed reporter deposition (CARD) technique to enhance staining results for use in automated true colour image analysis, to assess the reproducibility of systematic tissue sampling in the angiogenic hot spot selection, and quantification of microvessel density (MVD) and other vessel characteristics. The latter data were compared between six metastasised tongue squamous cell carcinomas, vs. four non-metastasised. In comparison to the standard immunohistochemical protocol with anti-CD34 antibodies, CARD amplification resulted in both more intensely stained and larger numbers of vessels. Averaging the 10 most vascularised fields of the 40 to 60 systematically sampled fields in a tissue section resulted in an overall acceptable interobserver reproducibility for most assessed vessel parameters (r> or =0.76 and p< or =0.01). The percentage vessels with diameter