Comparison of three staining methods to identify eosinophils in formalin-fixed canine skin

作者：Gomes Paulo; Torres Sheila M F^*; Plager Douglas A; Jessen Carl R; Lee James J

来源：Veterinary Dermatology, 2013, 24(3): 323-+.

DOI：10.1111/vde.12029

Background - Studies comparing the ability of staining methods to detect eosinophils in formalin-fixed canine skin are lacking.
Hypothesis/Objectives - The aim of this study was to compare the effectiveness of eosinophil peroxidase monoclonal antibody (EPXmAb), Luna and haematoxylin and eosin (H&E) to detect eosinophils in fixed canine skin by assessing the following parameters: (i) specificity of eosinophil staining; (ii) extracellular eosinophil granule staining; (iii) tissue background staining; (iv) contrast between eosinophil and surrounding tissue staining; and (v) differences in the number of eosinophils detected by each stain.
Methods - Three serial sections of formalin-fixed, paraffin-embedded tissues of predominantly eosinophilic skin diseases (n = 8), noneosinophilic skin diseases (n = 7) and normal canine skin (n = 1) were stained with the three stains. Each parameter was independently assessed and scored by two investigators.
Results - Luna and EPXmAb were specific in detecting eosinophils. The EPXmAb was significantly more effective than Luna (P < 0.001) and H&E (P < 0.001) in its ability to detect extracellular eosinophil granules (i.e. to detect intact and released eosinophil granules). The EPXmAb showed significantly less background staining compared with Luna (P = 0.0005). Moreover, significantly more stain contrast was noted with EPXmAb compared with Luna (P = 0.003) and H&E (P < 0.001). There was no significant difference between the mean eosinophil counts among the three stains.
Conclusion and clinical importance - The three stains were shown to be useful to detect and quantify eosinophils in fixed canine skin. The EPXmAb-based immunohistochemical stain proved to be a novel tool to detect eosinophils in canine skin.

• 出版日期2013-6