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Xylene Free Dewaxing

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Mel John del Barrio Aotea Pathology

A.k.a Deparaffinization A key point in the histology staining protocols Dewaxing agents be miscible with alcohol, wax and SAFE!!!

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

Melts latex gloves Melts the soles of your shoes!!! CNS depression Skin, eye irritant Nephrotoxic/Hepatoxic Teratogen/Fetotoxic Bone Marrow suppressant Carcinogen?

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

Is a detergent used to assist in dishwashing Cheap As!!! Readily available

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

Used for cleaning Laboratory Utensils, Benches etc Used to remove LCS (Ventana) from IHC slides Dewaxing of Slides for Staining

1st published in Laboratory Investigation journal in 2001 Sweden and USA A popular topic in Histonet

Place slides in oven for 5 to 10 minutes @ 60 C

Place slides in waterbath heated at 70C for 1 min and repeat

Wash slides in running hot tap water

Stain Slides

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

All slides (normal and soapy) were rated between 3 to 5 Soapy slides where less superior Diagnosis is not affected Difficulty of distinguishing

Specimen Appendix

KW

RM

JM

Average

5 Appendix (S) Bowel 5 Bowel(S) 4 Breast 4 Breast (S) 5 Liver 5 Liver(S) 4 Skin 5 Skin(S) 4 4

3 3 4 3 5 3 4 3 5 4

5 5 5 4 5 5 5 5 5 4

4.33 4.00 4.67 3.67 4.67 4.33 4.67 4.00 5.00 4.00

Dewaxing with DWS is cheaper to use The health hazards of DWS is lesser H & E staining times will be shorter

THANK YOU!!!

Dr. Karen Wood Dr. John Maccaferty Dr. Rupa Maitra Kerry Patience Abed Kader Rene Buesa Leanne Ward Kim Brown Emily Workman

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 Mller.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

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