Gobbo Francesca, Sarli Giuseppe, De Silva Margherita, Galiazzo Giorgia, Chiocchetti Roberto, Morini Maria
Department of Veterinary Medical Sciences, University of Bologna, Ozzano dell'Emilia, 40064 Bologna, Italy.
Vet Sci. 2021 Oct 12;8(10):229. doi: 10.3390/vetsci8100229.
Immunohistochemistry (IHC) is a widely used technique in diagnostic pathology, but the simultaneous analysis of more than one antibody at a time with different chromogens is rather complex, time-consuming, and quite expensive. In order to facilitate the identification of mast cells (MCs) during immunohistochemical analysis of membrane and/or nuclear markers, we propose a new staining method that includes the association of IHC and toluidine blue as a counterstain. To achieve this goal, we tested c-kit, Ki67, and cannabinoid receptor 2 on several cases of cutaneous canine mast cell tumors (MCTs), cutaneous mastocytosis, and atopic dermatitis. The results obtained show how this double staining technique, although limited to non-cytoplasmic markers and of little use in poorly differentiated MCTs in which MC metachromasia is hard to see, can be used during the evaluation of nuclear and/or membranous immunohistochemical markers in all canine cutaneous disorders, especially if characterized by the presence of a low number of MCs. It can help to evaluate those MCTs in which neoplastic MCs must be clearly distinguished from inflammatory cells that can infiltrate the tumor itself, in facilitating the calculation of the Ki67 index. Moreover, it can be used to study the expression of new markers in both animal and human tissues containing MCs and in MC disorders.
免疫组织化学（IHC）是诊断病理学中广泛使用的技术，但同时使用不同显色剂对多种抗体进行分析相当复杂、耗时且成本高昂。为了在对膜和/或核标记物进行免疫组织化学分析时便于识别肥大细胞（MCs），我们提出了一种新的染色方法，该方法包括将免疫组织化学与甲苯胺蓝作为复染剂相结合。为实现这一目标，我们在几例犬皮肤肥大细胞瘤（MCTs）、皮肤肥大细胞增多症和特应性皮炎病例中检测了c-kit、Ki67和大麻素受体2。获得的结果表明，这种双重染色技术虽然仅限于非细胞质标记物，且在难以观察到MC异染性的低分化MCTs中作用不大，但可用于评估所有犬皮肤疾病中的核和/或膜免疫组织化学标记物，特别是在MC数量较少的情况下。它有助于评估那些必须将肿瘤性MC与可浸润肿瘤本身的炎性细胞清楚区分的MCTs，有助于计算Ki67指数。此外，它可用于研究含有MCs的动物和人类组织以及MC疾病中新标记物的表达。
