Determining tartrate-resistant acid phosphatase in decalcified bone samples pig Sus domesticus

SUMMARYIntroduction: Nowadays, Immunohistochemistryare one of the most importanttechniques in the histopathological diagnosis.Despite of handling and processing cautions,both of them may generate changesin the protein structure, masking antigens,preventing antibody identification. Amongdifferent antigen retrieval methods proposedby the literature, the humid heat withSteamer has been widely accepted for itssimplicity, control and low cost.Objective: In this paper a protocol forhumid heat antigen retrieval use in bonetissue samples that had been previouslyfixed in formaldehyde, decalcified withethylenediaminetetraacetic (EDTA ), andembedded in paraffin is proposed.Materials and methods: Samples of pig’sbone that had been prepared with conventionaltechniques, from the collection ofthe pathology laboratory of the Universityof Valle were used. New fixed process wasperformed in glutaraldehyde and a steamerwas used to enhance antigen exposure. Theanti-TRAP DEKOs ®, mouse monoclonalantibody was used as primary antibody,and the KIT UltraVision LP Large VolumeDetection System HRP Polymer ® wasused as a secondary antibody.Results: In all samples exposed to primaryantibody in 1:20 and 1:40 dilutions immunostainingmononuclear cells compatiblewith preosteoclasts were observed; immunostainingmultinucleated giant cellsconsistent with osteoclasts were described.Conclusions: Humid heat for antigenicrecovery is a reliable method for the recoveryof bone antigen samples fixed withformaldehyde.