Clarified Butter as Xylene Substitute in Routine Histopathology of Oral and Maxillofacial Biopsies

ABSTRACT

Introduction: An ideal clearing agent should remove alcohol from the tissue specimens to enable the penetration of paraffin wax which provides sturdiness to the tissue specimen. Xylene has toxic effects and is still used because of its accurate clearing property and maintaining the staining quality. Natural alternatives to xylene provide a less toxic environment for histopathological technicians. Aim: The study is to evaluate the accuracy of clarified butter as xylene substitute in routine histopathological tissue processing procedure. Materials and Methods: 40 paired soft tissue specimens were obtained from the Department of Oral pathology and microbiology and subjected to routine histopathological tissue processing with xylene and ghee as clearing agents. Post processing, the tissue specimens were immersed in paraffin wax overnight, embedded, sectioned and stained using hematoxylin and eosin. Histopathological criteria namely nuclear staining, cytoplasmic staining, artefacts and background staining were evaluated and the scores were tabulated and statistical analysis was done using SPSS version 20 by IBM. Results: Our study results showed that tissue specimens processed in xylene and ghee had similar nuclear staining. In cytoplasmic staining, tissue specimens processed in ghee had superior results than xylene processing. Artefacts were present more in xylene processing than ghee processing. Background staining was more appreciated in xylene processed tissue specimens than ghee processing. Conclusion: Substituting xylene with ghee showed better results as a clearing agent in routine histopathological tissue processing. Ghee as a clearing agent is non toxic, cost effective compared to xylene and a safer alternative.