OHS026
Safe Work Procedure
Faculty/Division
Medicine
Document number
SOMS GBD SWP 032
School/ Divisional Unit
School of Medical Science
Initial Issue date
13-03-2009
Current version
1.0
Current Version
Issue date
20-04-2010
Next review date
20-04-2013
The Writing Safe Work Procedures Guideline (OHS027) should be consulted to assist in the completion of this
form.
Safe Work Procedure Title and basic description
Title:
Haematoxylin and Eosin (H&E) staining of frozen muscle sections
Description: Staining of frozen muscle tissues for histological analyses
Associated risk assessment title and location: RA handle human tissue 020, RA Haematoxylin and Eosin staining 005, RA waste
management 015 located in room 209B.
Describe the activity or process
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Air dry cut muscle sections at room temperature.
Place slides into 1xPBS solution for 5 minutes.
Place slides into water for 5 minutes.
Place slides into Harris Haematoxylin for 3-5 minutes.
Gently wash in running tap water for 3 minutes.
Place slides into acid alcohol for 30 seconds.
Place slides into Scotts bluing solution for 60 seconds.
Wash in running tap water for 3 minutes.
Place sections in Eosin for 2 minutes.
Rinse in tap water.
Place slides into 70% ethanol for 2 minutes.
Place slides in 95% ethanol for 2 minutes.
Place slides in 100% absolute ethanol for 2 minutes.
Place slides into a second 100% absolute alcohol for 2 minutes.
Place slides into xylene for 2 minutes (in fume hood).
Placed slides into a second xylene for 2 minutes (in fume hood).
Place slides into a third xylene for 2 minutes (in fume hood).
Remove slides from the slide holder one at a time and mount with with DePex mounting.
Leave slides in the fume hood to dry.
Store stained slides in a slide container.
List all resources required including plant,
chemicals, personal protective clothing and
equipment, etc
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PPE
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1% Alcoholic Eosin (Fronine laboratory supplies)
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70% ethanol
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95% ethanol
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Absolute ethanol
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Acid Alcohol
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DePex mounting medium (BDH laboratory supplies)
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Fume Hood
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Glass slides
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Glass slides holder
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Harris Haematoxylin
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xylene
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PBS
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Safety glasses
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Scotts water
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Page 1 of 2
Safe Work Procedure
Date Effective: 01/01/2007
Uncontrolled document when printed
Current Version: 1.2, 15/08/2007
List potential hazards and risk controls including
specific precautions required
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Potential Chemical and dangerous goods damage
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Ethanol – Highly flammable. Harmful. Cause central nervous system damage, eye, skin and respiratory tract irritation,
gastrointestinal irritation. Use gloves and safety goggles when preparing solutions in a well-ventilated area.
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DePex and Xylene –Flammable. Harmful by inhalation and in contact with skin and eyes. Causes irritation. Use
gloves, laboratory coat and work in a fume hood.
Potential human sample contamination – all users have been immunized against Hepatitis B, all samples are considered to be
infectious and appropriate PPE is always used, instruments used during procedure are soaked in 70% alcohol, then rinsed with
distilled water after using.
Cuts from broken slides – May cause skin cuts if broken. Use gloves when handling unbroken slides and tweezers if broken.
Always handle glass slides with care.
List emergency shutdown instructions
List clean up and waste disposal requirements
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Dispose gloves, tips, tubes in biological waste or autoclaved waste bin (human tissue experiment)
Dispose all chemical waste in chemical waste bin
Dispose leftover slides in sharp bin
List legislation, standards and codes of practice used
in the development of the SWP
NSW OHS Act 2000
NSW OHS Regulation 2001
Australia Dangerous Goods Code
Code of Practice for the Labelling of Workplace Substances
AS/NZS 2243.2:2006. Safety in laboratories. Part 2: Chemical aspects
AS/NZS 2243.6-1990. Safety in laboratories. Part 6: Mechanical Aspects.
AS/NZS 2243.3:2006 Safety in Laboratories Part 7 Electrical aspects
AS/NZS 2161.1:2000 Occupational Protective Gloves – Selection, Use and Maintenance
AS/NZS 1336:1997 Recommended Practices for Occupational Eye Protection
Supervisory approval, training, and review
Supervisor: Prof E Burcher
Signature:
Plant custodian:
Signature
List competency required – qualifications, certificates, licencing, training - eg course or instruction:
SWP review date: 20-04-2013
Responsibility for SWP review: Fei Shang
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Page 2 of 2
Safe Work Procedure
Date Effective: 01/01/2007
Uncontrolled document when printed
Current Version: 1.2, 15/08/2007