



ORIGINAL ARTICLE Year : 2012 | Volume : 4 | Issue : 1 | Page : 37-42

Liquid dish washing soap: An excellent substitute for xylene and alcohol in hematoxylin and eosin staining procedure



Surekha Ramulu1, Anila Koneru1, Shamala Ravikumar1, Priyadarshini Sharma2, D. N. S. V Ramesh3, Ramesh Patil4

1 Department of Oral Pathology, Navodaya Dental College, Raichur, Karnataka, India

2 Department of Periodontology, Navodaya Dental College, Raichur, Karnataka, India

3 Department of Oral Medicine and Radiology, Navodaya Dental College, Raichur, Karnataka, India

4 Department of Community Medicine, Navodaya Dental College, Raichur, Karnataka, India



Date of Web Publication 10-Sep-2012

Correspondence Address:

Surekha Ramulu

Department of Oral Pathology, Navodaya Dental College, Raichur, Karnataka

India

Source of Support: None, Conflict of Interest: None Check

DOI: 10.4103/0975-8844.99890



Abstract

Aims: Liquid dish washing solution (DWS) was used as a substitute for xylene to dewax tissue sections during hematoxylin and eosin (H and E) staining. The aim was to test and compare the hypothesis that xylene-ethanol free (XEF) sections deparaffinized with diluted DWS are better than or at par with the conventional H and E sections. Materials and Methods: Fifty paraffin-embedded tissue blocks was included. One section was stained with conventional HandE (group A) and the other with XEF HandE (group B) staining method. Slides were scored for parameters: nuclear, cytoplasmic, clarity, uniformity, and crispness of staining. Z test was used for statistical analysis. For accuracy of diagnosis, sensitivity, specificity, positive predictive value, and negative predictive value were tested. Results: Adequate nuclear staining was noted in 94% in group A and 96% in group B, ­adequate cytoplasmic staining in 92% in group A and 86% in group B, clarity in 94% of group A and 96% of group B sections, uniform staining in 92% of group A and 80% of group B sections, crisp stain in 96% of group A and 88% of group B sections, and 94% of group A sections stained adequately for diagnosis as compared with 90% in group B sections. Conclusion: Liquid DWS can be used as an alternative and effective substitute to xylene and ethanol in routine HandE staining procedure.

Keywords: Alcohol, dewaxing, eosin, hematoxylin, liquid dish washing soap, xylene

How to cite this article:

Ramulu S, Koneru A, Ravikumar S, Sharma P, Ramesh D, Patil R. Liquid dish washing soap: An excellent substitute for xylene and alcohol in hematoxylin and eosin staining procedure. J Orofac Sci 2012;4:37-42

How to cite this URL:

Ramulu S, Koneru A, Ravikumar S, Sharma P, Ramesh D, Patil R. Liquid dish washing soap: An excellent substitute for xylene and alcohol in hematoxylin and eosin staining procedure. J Orofac Sci [serial online] 2012 [cited 2020 Sep 23];4:37-42. Available from: http://www.jofs.in/text.asp?2012/4/1/37/99890

Introduction

To test the hypothesis that xylene-ethanol free (XEF) sections deparaffinized with diluted DWS are better than or at par with the conventional H and E section. To compare the efficacy of XEF sections with that of conventional H and E sections in producing adequate H and E staining for diagnosis. To produce XEF histologic sections equivalent to conventional ones by using liquid DWS and Harri's hematoxylin stain instead of Mayer's hematoxylin stain so as to device an optimal staining procedure that would be simple, safe, cost- and time-effective.

Materials and Methods

Table 1: Conventional H and E staining procedure



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Table 2: Xylene and alcohol-free H and E staining procedure



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Nuclear staining (adequate = score 1, inadequate = score 0) Cytoplasmic staining (adequate = score 1, inadequate = score 0) Clarity of staining (present = score 1, absent = score 0) Uniformity of staining (present = score 1, absent = score 0) and Crispness of staining (present = score 1, absent = score 0)

Results

Adequate nuclear staining was noted in 94% of group A as compared with 96% of group B ( P > 0.05).

> 0.05). When cytoplasmic staining was observed, 92% of group A showed adequate cytoplasmic staining as compared with 86% of group B ( P > 0.05).

> 0.05). Clarity was present in 94% of group A and in 96% of group B sections ( P > 0.05).

> 0.05). Uniform staining was present in 92% of group A and 80% of group B sections ( P > 0.05).

> 0.05). Crisp stain was noted in 96% of group A and 88% of group B sections ( P > 0.05) [Table 3].

> 0.05) [Table 3]. Table 3: Staining pattern in group A and group B



Click here to view The staining was found to be adequate for diagnosis in 94% of group A sections as compared with 90% of group B sections ( P > 0.05) [Table 4].

> 0.05) [Table 4]. Table 4: Scores for the adequacy for diagnosis of stained sections in groups A and B



Click here to view When compared between the two groups, 100% sensitivity and 60% specificity was observed.

The PPV and NPV were found to be 95.7% and 100%, respectively.

Chi-square value = 19.07 (P < 0.0005), which was highly significant [Table 5]. Table 5: Sensitivity, specificity, positive predictive value, and negative predictive value for accuracy of diagnosis



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Discussion

Figure 1: Photomicrographs showing adequacy and clarity of staining in epithelial tissue (a) conventional H and E (20×) and (b) XEF H and E staining (20×); and photomicrographs showing adequacy and clarity of nuclear and cytoplasmic staining (c) conventional H and E (40×) and (d) XEF HandE staining (40×)



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Figure 2: Photomicrographs showing (a and b) out-of-focus areas in XEF H and E staining (40×)



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Figure 3: Photomicrographs showing crisp staining in salivary glands and muscle: (a) Salivary gland with conventional HandE (20×), (b) salivary gland with XEF H and E staining (20×), (c) muscle tissue with conventional HandE (40×), (d) muscle tissue with XEF H and E staining (40×)



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Figure 4: Photomicrographs showing crisp staining in cartilage and adipose tissue: (a) Cartilage with conventional HandE (20×), (b) cartilage with XEF HandE staining (20×), (c) adipose tissue with conventional HandE (40×), (d) adipose tissue with XEF HandE staining (40×)



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Conclusions

Liquid DWS can be used as an alternative and effective substitute to xylene and ethanol in routine H and E staining procedure. Harris hematoxylin can be used instead of Mayer's with similar nuclear staining pattern in XEF procedure. Using liquid DWS as a substitute for xylene and alcohol gives an added advantage of it being eco-friendly, non-toxic, non-inflammable, and cost- and time-effective.

Acknowledgments

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