Histochemical and immunohistochemical evaluation of mouse skin histology: comparison of fixation with neutral buffered formalin and alcoholic formalin

Abstract

Histological analysis is a cornerstone of skin disease diagnosis, and the stratified nature of the skin presents many technical problems in the preparation of sections of acceptable quality. Furthermore, histological artifacts may be exacerbated in pathological states that compromise cutaneous integrity. Therefore, great care must be taken when selecting the conditions, particularly fixative type, to ensure that meaningful conclusions may be drawn. The goal of this work was to compare the morphology of mouse skin as revealed using a range of histological stains, and the antigenicity of key cutaneous proteins using immunohistochemistry (IHC) following fixation in either 10% neutral buffered formalin (NBF) or alcoholic formalin (AF). Although 10% NBF fixation is routinely used for histology, providing good retention of morphology and structure for special staining, AF fixation more effectively maintained antigenicity for the cutaneous markers investigated in this study. A comprehensive assessment of the two fixatives and histological techniques to optimize morphology, structure, and antigenicity in the analysis of mouse skin is presented.

Keywords:

• Fixation
• Histology
• Immunohistochemistry
• Skin
• Special stains