Abstract

Objectives: Bone-decalsification is still a time consuming and laboring process in histopathology laboratories. In this study, we have aimed a comparision of decalsification degrees and staining properties of compact bone tissue decalcificated by formic acid, Biodec-R, and Decalcifier II as decalcification agents.

Materials and Methodology: A total of 6 healthy male rats (200-220 g) were used in this study. Rats were decapitated by cervical dislocation. Femurs were removed and 0.5 cm long pieces from these femurs were fixed in 10% formaldehyde for 36 hours. Subsequently, the bone-tissues were stored in decalcification fluids at room temperature for six days. The bone-tissue samples were processed by routine tissue procedures. They were further processed for light microscopic examination and stained with hematoxylin-eosin, Gomori’s trichrome, and Periodic acid-Schiff. We have examined the bone sections under a Leica DFC280 light microscope and Leica Q Win Image Analysis System (Leica Micros Imaging Solutions Ltd.; Cambridge, U.K).

Results: When all three decalcification agents were applied for equal time periods and at the same experimental design, it was observed that formic acid is more effectible for the preservation of natural structure of the bone tissue and on the quality of the staining properties. It was observed that Biodec R and Decalcifier II are similar to each other in terms of staining properties and preservation of structural details of cells and tissue. 

Conclusion: Formic acid decalcification is adviced for histologic staining and for a higher qualitiy of microscopic view during histological examination of compact bone tissues.

Key Words: Bone; Decalcification; Formic Acid; Biodec-R; Decalcifier II.