Staining of mast cells (MCs), together with porcine ones, is critically dependent upon the fixation and marking method. Within the pig, mucosal and submucosal MCs don’t stain or stain solely faintly after formalin fixation. Some fixation strategies are significantly really helpful for MC staining, for instance the fixation with Carnoy or lead salts. Zinc salt fixation (ZSF) has been reported to work excellently for the preservation of fixation-sensitive antigens. The purpose of this examine was to ascertain a dependable histological methodology for counting of MCs within the porcine intestinum.
For this goal, completely different tissue fixation and marking strategies that additionally enable potential subsequent immunohistochemical investigations have been evaluated within the porcine mucosa, in addition to submucosa of small and huge gut. Tissues have been fastened in Carnoy, lead acetate, lead nitrate, Zamboni and ZSF and stained subsequently with both polychromatic methylene blue, alcian blue or toluidine blue.
For the primary time our examine reveals that ZSF, a heavy metallic fixative, preserves metachromatic staining of porcine MCs. Zamboni fixation was not appropriate for histochemical visualization of MCs within the pig gut. All different examined fixatives have been appropriate. Alcian blue and toluidine blue co-stained intestinal goblet cells which made a prima facie identification of MCs tough.
The polychromatic methylene blue proved to be the optimum staining. So as to examine MC counting outcomes of the completely different fixation strategies, tissue shrinkage was taken into consideration. As even the identical fixation induced shrinkage-differences between tissue from small and huge gut, various factors for every single fixation and intestinal localization needed to be calculated.
Tissue shrinkage various between 19% and 57%, the best tissue shrinkage was discovered after fixation with ZSF within the massive gut, the bottom one within the small gut after lead acetate fixation. Our examine emphasizes that MC counting outcomes from information utilizing completely different fixation strategies can solely be in contrast if the respective study-immanent shrinkage issue has been decided and quantification outcomes are adjusted accordingly.