Assessment of the Effectiveness of Xylene-free Haematoxylin and Eosin Staining versus the Conventional Approach: A Comparative Observational Study

Abstract

Introduction: Xylene, a commonly used hydrocarbon in histopathological laboratories, poses significant risks, including carcinogenicity. Identifying safer alternatives is essential for laboratory safety and environmental sustainability. Aim: To evaluate the feasibility and effectiveness of xylene-free Haematoxylin and Eosin (H&E) staining as a safer and more environmentally friendly alternative to conventional xylene-based staining methods. Materials and Methods: A comparative observational study was conducted in the Department of Pathology at Karpaga Vinayaga Institute of Medical Sciences and Research Centre (KIMS & RC), Chengalpattu, Tamil Nadu, India, over a duration of three months (January 2024 to March 2024). The study included 50 specimen blocks (100 slides), with two sections taken from each block: group A (conventional staining) and group B (xylene-free staining). The 1.7% dishwashing liquid solution used in the xylene-free method was prepared by mixing 25 mL of commercially available dishwashing solution with 1,500 mL of distilled water. The staining quality, cellular architecture and overall staining effectiveness were analysed statistically using Statistical Package for Social Sciences (SPSS) version 25.0. Results: The analysis shows that 49 (98%) of the slides in group A and 36 (72%) of the slides in group B had distinct nuclei and cytoplasm. The analysis of staining quality indicated that 45 (90%) of group A slides had good quality, whereas only 25 (50%) of group B slides achieved this standard. Overall quality was satisfactory in 49 (98%) of group A slides and only 36 (72%) of group B slides. Conclusion: Xylene-free H&E staining is a viable alternative with comparable diagnostic quality and improved safety and environmental benefits. However, slight reductions in staining clarity suggest a need for further optimisation. Future research should focus on improving deparaffinisation efficiency and refining methodologies to enhance nuclear and cytoplasmic clarity while maintaining safety and sustainability benefits.

Keywords

• deparaffinisation
• histological techniques
• staining and labelling