Abstract
Background: Deparaffinization of wax sections in histopathology requires Xylene which is a harmful and toxic compound. It has been successfully replaced by many common household products such as dishwashing liquid, soaps and mineral oils. However, all these substitutes require higher temperatures of upto 90oC for effective deparaffinization. Aim: To evaluate the temperature dependent efficacy of dishwashing liquid in deparaffinization of histopathological sections as a substitute to the routine toxic Xylene. Method: 100 tissue sections deparaffinised by conventional Xylene and stained by Hematoxylin and Eosin method were objectively scored against 300 matched sections deparaffinised using dilute dishwashing liquid method at 65oC/75oC/90oC (100 each) by a single blinded pathologist. Results: Based on Wilcoxon signed ranks test, at 90oC the Xylene free method stood on par with the conventional method (p=0.422, p>0.05) proving Xylene free sections were as good or better than the matching conventional sections in 76% of the comparisons. Xylene free sections at 65oC and 75oC were found to be inferior to the conventional method in 80% and 69% of the comparisons respectively and also statistically significant from the conventional method (p=0.002 and 0.019 respectively). Conclusion: The new Xylene free method using dishwashing liquid is highly temperature sensitive and effective only at higher temperatures.
Highlights
The backbone of daily histopathological diagnostic work is the paraffin section usually stained with Hematoxylin and Eosin
Any substitute that minimizes the use of Xylene in experiments, reduces tissue staining time and does not compromise its quality will be efficient for diagnostic reasons and valuable for maintaining a safe laboratory environment
The Xylene-free sections at 90oC were judged to be as good as or better than the matching conventional sections in 76% of comparisons and poorer in 24% of the comparisons, which is analogous to results obtained by Falkholm et al [2]
Summary
The process of deparaffinisation of the slides using Xylene is an important preliminary step before the staining process,which makes the tissue sections to uptake the Haemotxylin and Eosin stain properly. This makes Xylene an inevitable compound in histopathology due to its paraffin solvent action. Deparaffinization of wax sections in histopathology requires Xylene which is a harmful and toxic compound It has been successfully replaced by many common household products such as dishwashing liquid, soaps and mineral oils. All these substitutes require higher temperatures of upto 90oC for effective deparaffinization. Conclusion: The new Xylene free method using dishwashing liquid is highly temperature sensitive and effective only at higher temperatures
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