Download PDF
CC BY-NC-ND 4.0 · J Lab Physicians 2019; 11(02): 118-122
DOI: 10.4103/JLP.JLP_169_18
Original Article

A substitute to xylene in deparaffinization and clearing prior to coverslipping in histopathology

Nasar Yousuf Alwahaibi
Department of Allied Health Sciences, College of Medicine and Health Sciences, Sultan Qaboos University, Muscat, Oman, India
,
Sirin Hamed Aldughaishi
Department of Allied Health Sciences, College of Medicine and Health Sciences, Sultan Qaboos University, Muscat, Oman, India
› Author Affiliations Financial support and sponsorship Nil
› Further Information
Also available at
  PDF Download Permissions and Reprints

Abstract

INTRODUCTION: Deparaffinization and clearing prior to coverslipping are important steps in all staining methods in histopathology. Xylene is the most commonly used agent worldwide. However, xylene is toxic. We evaluated safer alternative dewaxing and clearing agents prior to coverslipping in a histopathology laboratory.

MATERIALS AND METHODS: Thirteen different fresh surgical tissues were cut into two halves. One half processed using xylene and the other half processed using UltraClear™. Five groups were designed. For each Group of A, B, C, and D, 100 slides were cut from xylene-processed blocks. For Group E, 100 slides were cut from UltraClear™-processed blocks. Group A is the standard method. Group B evaluates UltraClear™ as a dewaxing agent only. Group C evaluates UltraClear™ as a clearing agent prior to coverslipping only. Group D evaluates UltraClear™ as both dewaxing and clearing agents prior to coverslipping. Group E evaluates UltraClear™ as both dewaxing and clearing agents prior to coverslipping. Six parameters were evaluated: nuclear staining, cytoplasmic staining, cell morphology, clarity of staining, uniformity of staining, and cost.

RESULTS: Groups B, C, and D showed 79% (P = 0.054), 83% (P = 0.221), and 80% (P = 0.079) adequacy when compared with Group A (89%), respectively. However, Group E showed only 76% (P = 0.016) adequacy. UltraClear™ is more expensive than xylene.

CONCLUSION: UltraClear™ is a promising dewaxing agent. It is also a good clearing agent for use prior to coverslipping in histopathology laboratory. Cost-benefit balance between safety of laboratory workers, good quality staining, and cost-effective strategy needs to be further studied.

Key words

Clearing agent prior to coverslipping - dewaxing agent - histopathology - UltraClear™ - xylene


Publication History

Received: 11 December 2018

Accepted: 22 March 2019

Article published online:
06 April 2020

© 2019.

Thieme Medical and Scientific Publishers Private Ltd.
A-12, Second Floor, Sector -2, NOIDA -201301, India

 

  • References

  • 1 Udonkang M, Eluwa M, Ekanem A, Sharma TB, Asuquo OR, Akpantah AO. Bleached palm oil as substitute for xylene in histology. J Pharm Clin Res 2014;8:8-17.
  • 2 Material Safety Data Sheet (MSDS) for Xylene. Fisher Scientific. Available from: https://www.fscimage.fishersci.com/msds/25150.htm. [Last accessed on 2018 Jul 20].
  • 3 Buesa RJ. Mineral oil: The best xylene substitute for tissue processing yet? J Histotechnol 2000;23:143-9.
  • 4 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.
  • 5 Material Safety Data Sheet (MSDS) for UltraClear™. Avantor Performance Materials B.V. Available from: http://www.avantormaterials.com/documents/MSDS/DEV/GB/MSDS_3905_GB.pdf. 2010. [Last accessed on 2018 Jul 15].
  • 6 Alwahaibi N, Aljaradi S, Alazri H. Alternative to xylene as a clearing agent in histopathology. J Lab Physicians 2018;10:189-93.
  • 7 Bancroft JD, Gamble M. Theory and Practice of Histological Techniques. 6th ed. Philadelphia, USA: Churchill Livingstone Elsevier; 2008. p. 126-7.
  • 8 Singh A, Hunasgi S, Koneru A, Vanishree M, Ramalu S, Manvikar V, et al. Comparison of honey with ethanol as an oral cytological fixative: A pilot study. J Cytol 2015;32:113-7.
  • 9 Pandey P, Dixit A, Tanwar A, Sharma A, Mittal S. A comparative study to evaluate liquid dish washing soap as an alternative to xylene and alcohol in deparaffinization and hematoxylin and eosin staining. J Lab Physicians 2014;6:84-90.
  • 10 Buesa RJ, Peshkov MV. Histology without xylene. Ann Diagn Pathol 2009;13:246-56.
  • 11 Kunhua W, Chuming F, Tao L, Yanmei Y, Xin Y, Xiaoming Z, et al. Anovel non-toxic xylene substitute (SBO) for histology. Afr J Tradit Complement Altern Med 2012;9:43-9.
  • 12 Indu S, Ramesh V, Indu PC, Prashad KV, Premalatha B, Ramadoss K, et al. Comparative efficacy of cedarwood oil and xylene in hematoxylin and eosin staining procedures: An experimental study. J Nat Sci Biol Med 2014;5:284-7.
  • 13 Bruun Rasmussen B, Norring Hjort K, Mellerup I, Sether G, Christensen N. Vegetable oils instead of xylene in tissue processing. APMIS 1992;100:827-31.
  • 14 Negi A, Puri A, Gupta R, Chauhan I, Nangia R, Sachdeva A, et al. Biosafe alternative to xylene: A comparative study. J Oral Maxillofac Pathol 2013;17:363-6.
  • 15 Metgud R, Astekar MS, Soni A, Naik S, Vanishree M. Conventional xylene and xylene-free methods for routine histopathological preparation of tissue sections. Biotech Histochem 2013;88:235-41.
  • 16 Ananthaneni A, Namala S, Guduru VS, Ramprasad VV, Ramisetty SD, Udayashankar U, et al. Efficacy of 1.5% dish washing solution and 95% lemon water in substituting perilous xylene as a deparaffinizing agent for routine H and E staining procedure: A short study. Scientifica (Cairo) 2014;2014:707310.