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CC BY-NC-ND 4.0 · J Lab Physicians 2014; 6(02): 084-090
DOI: 10.4103/0974-2727.141504
Original Article

A comparative study to evaluate liquid dish washing soap as an alternative to xylene and alcohol in deparaffinization and hematoxylin and eosin staining

Pinki Pandey
Department of Pathology, UP Rural Institute of Medical Sciences and Research, Saifai, Etawah, Uttar Pradesh, India
,
Alok Dixit
Department of Pharmacology, UP Rural Institute of Medical Sciences and Research, Saifai, Etawah, Uttar Pradesh, India
,
Aparna Tanwar
Department of Pathology, Maharishi Markandeshwar Institute of Medical Sciences and Research, Sultanpur, Kumarhatti, Solan, Himachal Pradesh, India
,
Anuradha Sharma
Department of Pathology, M M Institute of Medical Sciences and Research, M M University, Mullana, Ambala, Haryana, India
,
Sanjeev Mittal
Department of Pathology, M M Institute of Medical Sciences and Research, M M University, Mullana, Ambala, Haryana, India
› Author Affiliations Source of Support: Nil
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ABSTRACT

Introduction: Our study presents a new deparaffinizing and hematoxylin and eosin (H and E) staining method that involves the use of easily available, nontoxic and eco-friendly liquid diluted dish washing soap (DWS) by completely eliminating expensive and hazardous xylene and alcohol from deparaffinizing and rehydration prior to staining, staining and from dehydration prior to mounting. The aim was to evaluate and compare the quality of liquid DWS treated xylene and alcohol free (XAF) sections with that of the conventional H and E sections.

Materials and Methods: A total of 100 paraffin embedded tissue blocks from different tissues were included. From each tissue block, one section was stained with conventional H and E (normal sections) and the other with XAF H and E (soapy sections) staining method. Slides were scored using five parameters: Nuclear, cytoplasmic, clarity, uniformity, and crispness of staining. Z-test was used for statistical analysis.

Results: Soapy sections scored better for cytoplasmic (90%) and crisp staining (95%) with a statistically significant difference. Whereas for uniformity of staining, normal sections (88%) scored over soapy sections (72%) (Z = 2.82, P < 0.05). For nuclear (90%) and clarity of staining (90%) total scored favored soapy sections, but the difference was not statistically significant. About 84% normal sections stained adequately for diagnosis when compared with 86% in soapy sections (Z = 0.396, P > 0.05).

Conclusion: Liquid DWS is a safe and efficient alternative to xylene and alcohol in deparaffinization and routine H and E staining procedure. We are documenting this project that can be used as a model for other histology laboratories.

Keywords

Alcohol - deparaffinization - eosin - hematoxylin - liquid dish washing soap - xylene-free staining


Publication History

Article published online:
19 April 2020

© 2014.

Thieme Medical and Scientific Publishers Private Ltd.
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  • REFERENCES

  • 1 Cook HC. Origins of tinctorial methods in histology. J Clin Pathol 1997;50:716-20.
  • 2 Q Imaging Camera Application Notes. H and Estain tissue documentation. Available from: http://www.qimaging.com/support/pdfs/he_technote.pdf. [Last accessed on 2013 Jan 01].
  • 3 Falkeholm L, Grant CA, Magnusson A, Möller E. Xylene-free method for histological preparation: A multicentre evaluation. Lab Invest 2001;81:1213-21.
  • 4 Kandyala R, Raghavendra SP, Rajasekharan ST. Xylene: An overview of its health hazards and preventive measures. J Oral Maxillofac Pathol 2010;14:1-5.
  • 5 Buesa RJ, Peshkov MV. Histology without xylene. Ann Diagn Pathol 2009;13:246-56.
  • 6 Ankle MR, Joshi PS. 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. J Oral Maxillofac Pathol 2011;15:161-7.
  • 7 National Institute for Occupational Safety and Health (NIOSH) Criteria for a recommended standard: Occupational exposure to xylene 1975. Available from: http://www.cdc.gov/niosh/75-168.html. [Last accessed on 2013 Jan 01].
  • 8 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.
  • 9 Buesa RJ, Peshkov MV. Complete elimination of xylene in practice of a histology laboratory. Arkh Patol 2011;73:54-60.
  • 10 Available from: http://www.en.wikipedia.org/wiki/Detergent. [Last accessed on 2013 Jan 01].
  • 11 Technical Information Guide. Available from: http://www.technogreen.us/wp-content/uploads/2011/09/3_Dish-Soap2.pdf. [Last accessed on 2013 Jan 01].
  • 12 Mayer′s Hematoxylin Protocol. Available from: http://www.ihcworld.com/_protocols/counterstain_solutions/mayer-hematoxylin.htm. [Last accessed on 2012 Mar 23].
  • 13 Chen CY, He T, Mao XL, Friis TE, Qin RH, Jian YT. A novel xylene substitute for histotechnology and histochemistry. Biotech Histochem 2010;85:231-40.
  • 14 Buesa RJ. Mineral oil: The best xylene substitute for tissue processing yet? J Histotechnol 2000;23:143-9.
  • 15 Stevens A, Wilson I. The haematoxylins and eosin. In: Bancroft JD, Stevens A, editors. Theory and Practice of Histological Techniques. 4th ed., Ch. 6. New York: Churchill-Livingstone; 1996. p. 99-112.
  • 16 Gill G. H and E staining: Oversights and insights. Available from: http://www.dako.com/08066_12may10_webchapter 13.pdf. [Last accessed on 2013 Jan 01].
  • 17 Ellis R. Problems in histopathological technique. Available from: http://www.ihcworld.com/royellis/problems/problem25.htm. [Last accessed on 2013 Jan 01].
  • 18 Hamil JR, Spencer JM, Morgan MB. Histotechnique and staining troubleshooting. On Atlas of Mohs and Frozen Section Cutaneous Pathology. Ch. 20. New York: Springer Link; 2009. Available from: http://www.books.google.co.in/books. [Last accessed on 2013 Jan 01].