Cedarwood oil as an alternative to xylene as a clearing agent in histopathological tissue processing – A comparative study

Abstract

Background: Clearing in histopathological tissue processing should be able to make the tissues translucent and clear for the tissues to be visible under light microscopy and should render the clearing agent to be miscible with the dehydrant and the impregnation wax in the preceding and following processing steps. Xylene is a gold standard clearing agent but increasing concerns about the potential carcinogenicity, implementing eco-friendly agents in routine histopathology is necessary. Aim: The aim of the study is to assess the clearing ability of Cedarwood oil as an alternative to Xylene in routine tissue processing. Materials & Methods: The study was carried out in the Department of Oral Pathology and Microbiology. Formalin fixed 50 tissue samples of size 3-7mm were taken and subsequent dehydration done with acetone and alcohol. The dehydrated tissue is later processed using 90ml of Cedarwood oil with few drops of Xylene and Thymol. After clearing the tissues were subjected to impregnation and embedded in paraffin wax, later which sections were made and stained using H & E stain. Results: The results of our study on comparison showed better outcome in tissues processed with cedarwood oil than xylene. Statistical Significant correlation was observed in nuclear staining (p value = 0.001) ; cytoplasmic staining (p value = 0.08) and background staining (p value = 0.045) indicating a positive results on using cedarwood oil as clearing agent. Conclusion: The cedarwood oil can be considered as a safer natural alternative to xylene in laboratories. The cedarwood oil is eco - friendly and easily available with enhanced tissue processing qualities.

Preserving Stained Fish Specimens in Resin Epoxy

“…It was not for the entire body of an animal with pigmentation. [12] showed that many specimens were preserved with resin for a long time without harm, knowing that this study did not refer to whole living bodies such as fish or other animals. Whereas [13] showed that many plants and insects were preserved as living organisms in the natural resin known as amber without human intervention making these living organisms an important subject for genetic studies due to their excellent preservation.…”