Xylene Free Dewaxing
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Transcript of Xylene Free Dewaxing
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XYLENE FREE DEWAXING
Mel John del BarrioAotea Pathology
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Dewaxing
A.k.a “ Deparaffinization
A key point in the histology staining protocols
Dewaxing agents be miscible with alcohol, wax and SAFE!!!
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Xylene
A mixture of three aromatic hydrocarbons isomers related to benzene
102 synonyms Alkylation of crude oil,
Dehydrocyclodimerization 1st used in the 1950s as a safer
alternative Safety was questioned in the late 70s
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What Xylene Does?
Melts latex gloves Melts the soles of your shoes!!! CNS depression Skin, eye irritant Nephrotoxic/Hepatoxic Teratogen/Fetotoxic Bone Marrow suppressant Carcinogen?
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Information about the health effects of xylene led laboratory supplier to come up with a safer substitute that would still do the job of xylene.
Xylene substitutes is widely available.
Various journals published the use of other chemical for dewax
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Dishwashing soap
Is a detergent used to assist in dishwashing
Cheap As!!!
Readily available
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How does DWS clean your dishes
DWS contains a mixture of surfactants
Sodium Lauryl Sulfate
Surfactants lower surface tension of water
Surfactants molecules are composed of hydrophobic and water soluble component
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Sodium Lauryl Sulfate
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Dishwashing soap in Histology
Used for cleaning Laboratory Utensils, Benches etc
Used to remove LCS (Ventana) from IHC slides
Dewaxing of Slides for Staining
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Dewaxing with Soap
1st published in Laboratory Investigation journal in 2001
Sweden and USA A popular topic in Histonet
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H & E (Modified)
Place slides in oven for 5 to 10 minutes @ 60 C
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Place slides in waterbath heated at 70C for 1 min and repeat
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Wash slides in running hot tap water
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Stain Slides
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Appendix, Breast, Bowel, Liver and Skin were used
Slides were placed in Ordinary slides and Superfrost Plus
Modified H&E protocol done Slides were randomly labelled Slides were evaluated by 3 pathologists Slides were rated from 1=Very Poor to
5=Excellent
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Appendix
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Bowel
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Breast
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Liver
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Skin
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Findings
All slides (normal and soapy) were rated between 3 to 5
Soapy slides where less superior
Diagnosis is not affected
Difficulty of distinguishing
Specimen
KW RM JM Average
Appendix5 3 5 4.33
Appendix (S) 4 3 5 4.00
Bowel5 4 5 4.67
Bowel(S)4 3 4 3.67
Breast4 5 5 4.67
Breast (S) 5 3 5 4.33
Liver5 4 5 4.67
Liver(S)4 3 5 4.00
Skin5 5 5 5.00
Skin(S)4 4 4 4.00
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Conclusion
Dewaxing with DWS is cheaper to use
The health hazards of DWS is lesser
H & E staining times will be shorter
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THANK YOU!!!
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Acknowledgements
Dr. Karen Wood Dr. John Maccaferty Dr. Rupa Maitra Kerry Patience Abed Kader Rene Buesa Leanne Ward Kim Brown Emily Workman
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Reference Buesa RJ, Peshkov MV. Histology without xylene. Ann Diagn Pathol. 2009
Aug;13(4):246-56 Lars Falkeholm, Crawford A. Grant, Anders Magnusson, and Eva Möller.Xylene-Free
Method for Histological Preparation: AMulticentre Evaluation. LABORATORY INVESTIGATION. Vol. 81, No. 9, p. 1213, 2001
www.histonet.com http://www.osha.gov/SLTC/healthguidelines/xylene/recognition.html http://www.oehha.ca.gov/water/phg/pdf/xylen_c.pdf www.wikipedia.co.nz Ventana IHC staining manual http://chemistry.about.com/od/howthingswork/f/detergentfaq.htm www.chemistry.co.nz