Biofriendly substitutes for xylene in deparaffinization

Abstract

Background: Xylene is an aromatic hydrocarbon that is routinely used in histopathological laboratories. It is biohazardous and produces many toxic effects like carcinogenesis. A safer substitute for xylene is necessary to minimize its usage. Aim: The aim of this study was to evaluate the effectiveness of 1.7% dishwashing solution, 95% lemon water, and 100% coconut oil when compared to xylene as a deparaffinizing agent during hematoxylin and eosin (HE) staining. Materials and Methods: Fifteen paraffin-embedded tissue blocks were selected. Four sections were made from each block. One section was stained with conventional HE method using xylene (group A) as deparaffinizing agent and other three sections were stained with xylene-free HE method using 1.7% dishwashing solution (group B), 95% lemon water (group C), and 100% coconut oil (group D), respectively. Slides were scored blindly by a single pathologist considering the parameters such as nuclear and cytoplasmic staining; uniformity, clarity, and crispness of staining; and presence or absence of wax retention. Results: Adequate nuclear staining was noted in 100% of sections of groups A, B, C, and D (P 0.001), whereas adequate cytoplasmic staining was noted in 93.33% each in groups A, C, and D when compared with 100% in group B (P > 0.05). Uniform staining was present in 80% of groups A and B and in 73.33% of groups C and D (P > 0.05). Clarity of staining was present in 86.66% of groups A and B and in 80% of groups C and D (P > 0.05), whereas crispness of staining was seen in 73.33% of groups A and D, 86.66% of group B, and 80% of group C (P > 0.05). Wax retention was noted in 20% of groups A and B, and 26.66% of groups C and D (P > 0.05). Adequate staining for diagnosis was noted in 100% of group A sections followed by 93.33% in group B, 86.66% in group C, and 80% in group D as compared with 90% in group B (P > 0.05). Conclusion: Dishwashing solution, lemon water, and coconut oil can be used as safer and cost-effective substitutes to xylene for deparaffinization in HE staining procedure.