The Role of Fixation and Dimineralization on the Immunohistochemical Staining of an Anti Keratin Antibody and Bone Sialo Protein on CD-1 Mouse Tissue
The interaction between the epithelial root sheath (ERS) and the adjacent mesynchemal tissue is a process that is not yet fully understood. In order to study the interactions between these cells, one must have the optimal conditions to work with. This study was designed to create the optimal conditions, with regard to the choice of fIxative and demineralizer. Bone silo protein (BSP) and an anti keratin antibody (Keratin) were used to stain the cells along the epithelial root sheath of first molars of CD-1 mice at specific time points. Previously, when BSP was exposed to a Bouin's and Acid Formal Saline sample the cells along the epithelial root sheath provided positive staining. On the other hand those experiments with keratin provided less than consistent staining. Bouin's, Paraformeldahyde (PFA) and Ethanol (EtOH) are the three fixatives that were used in combination with the two demineralizers Acid Formal Saline (AFS) and ethylenediamine tetraacetic acid (EDTA). The result obtained demonstrated that Bouin's AFS as the lone fisative- demineralizer combination does not provide optimal conditions for staining. It did illustrate that the BSP would stain the ERS on the Bouin's AFS even though the keratin did not stain. Furthermore, the combination of EtOH and EDTA proved to be the most successful for the staining of the keratin, while no BSP was present. In addition, none of the fixative-demineralizer combinations stained for both BSP and Keratin; illustrating that, at least for BSP and Keratin, no fixative-demineralizer can be used alone.