A study to evaluate the efficacy of xylene-free hematoxylin and eosin staining procedure as compared to the conventional hematoxylin and eosin staining: An experimental study

Abstract

Context:Use of diluted dish washing solution (DWS) has been experimented successfully as a substitute for xylene to deparaffinize tissue sections during hematoxylin and eosin (H and E) staining.Aims:(1) Test the hypothesis that xylene- and methanol-free sections (XMF) deparaffinized with diluted DWS are better than or at par with conventional H and E sections. (2) To compare the efficacy of xylene-free sections with the conventional H and E sections.Settings and Design:Single blinded experimental study.Materials and Methods:Sixty paraffin blocks were considered. One section was stained with conventional H and E method (Group A) and the other with XMF H and E (Group B). Slides were scored for parameters; nuclear staining, cytoplasmic staining (adequate = score1, inadequate = score0), uniformity, clarity, crispness (present = score1, absent = score0). Score >/= 2 was inadequate for diagnosis and 3-5 was adequate for diagnosis.Statistical analysis used:Z test.Results:Adequate nuclear staining, 96.66% sections in group A and 98.33% in Group B (Z = 0.59, P>0.05); adequate cytoplasmic staining, 93.33% in group A and 83.33% in Group B (Z = 1.97, P<0.05); uniform staining, 70% in group A, 50% in group B (Z = 1.94, P<0.05), clarity present in 85% of group A, 88.33% of group B sections (Z = 0.27, P>0.05), crisp staining in 76.66% in group A and 83.33% in Group B (Z = 1.98, P<0.05), 88.33% Group A sections stained adequately for diagnosis as compared with 90% in Group B (Z = 0.17, P>0.05).Conclusion:Xylene- and methanol-free H and E staining is a better alternative to the conventional H and E staining procedure.

A Study of Xylene Free Hematoxylin and Eosin Staining Procedure

“…But the xylene free group of stained sections scored poorly in uniformity of staining and adequate cytoplasmic staining. [6] Ramulu et al in their study on liquid dish washing soap as a substitute for xylene and alcohol observed that liquid dish washing soap can be used as an alternative and effective substitute to xylene and alcohol in the routine H and E staining procedure. They noted that sections stained with xylene free method demonstrated adequate nuclear staining and better clarity of sections compared to the conventional…”

“…The ease of availability of these detergents with its non toxic nature makes it an attractive and cost effective alternative to xylene. [5,6,7,10] The present study intends to utilize the commercially available liquid dishwashing solution as a substitute for xylene and alcohol in dewaxing and staining.…”

“…Various alternatives have been tried to eliminate these toxic substances which are helpful both to the technician and to the environment. [5][6][7][8][9] These methods have met with varying degree of success.…”

Ecofriendly, Economic Surrogative for Xylene

“…The concentration of these components are well analysed by the manufacturer. 9 In this study, we have used dish wash liquid and the detergent liquid in a dilution of 1.5%,hence there are very minimal chances of toxicity to the laboratory personnel.…”

Microscopic Validation of Macroscopic In Vivo Images Enabled by Same-Slide Optical and Nuclear Fusion

“…Clinically, histopathologic staining with hematoxylin–eosin (HE) is the gold standard for identification of specific cells and tissues (6); however, ambiguity is common. To ensure proper identification, one or more immunomarkers are typically used, and precipitating pigments are used in conjunction with HE to label cells of interest (3,7).…”