Novel polyvinyl alcohol (PVA) based cryoprotection method that facilitates cutting frozen sections of decalcified human trabecular bone

Abstract

Processing adult human trabecular bone to obtain tissue sections suitable for research or diagnostic purposes has always been challenging, particularly in the preparation of adult bone specimens for advanced immunohistochemistry applications. In contrast to the majority of soft tissues, decalcified bone samples perform poorly under standard paraffin embedding techniques and immunolabeling protocols fail frequently, due to the loss of protein antigenicity observed. We report on a new, PVA based infiltration method that avoids excessive heat exposure to tissue samples during embedding. The developed PVA based infiltration medium provides sufficient structural support to the heterogenic morphology and distinct architecture of subchondral trabecular bone and adjacent articular cartilage. Furthermore, the addition of bovine serum albumin (BSA) to this infiltration solution guaranteed safe attachment of cryosections to glass slides. The protocol allows the preparation of high quality sections of adult human trabecular bone tissues which can be used for both classical histochemical stains and for immunohistochemistry, since protein antigenicity is satisfactorily preserved.