OHS026
Safe Work Procedure
Faculty/Division
School/ Divisional Unit
SCHOOL OF MEDICAL SCIENCES
MEDICINE
Document number
Initial Issue date
28-4-10
Current version
Current Version
Issue date
Next review date
28-4-13
SOMS GBD SWP
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: In Situ Hybridization
Description: Hybridization of DIG labelled probe to cellular mRNA.
Associated risk assessment title and location:
Describe the activity or process
- Cut paraffin embedded or frozen blocks into 8-10umthick tissue sections and mount on gelatine
coated slides or “Yellow Starfrost” slides.
- Dry in 500C oven for 1hr.
- Fix in ice cold 4% Paramormaldehyde, digest with Proteinase K, wash in PBS/Glycine.
- Pre-hybridize in hybridization buffer for 1-2hr at 38oC in humidified oven.
- Hybridize with DIG labelled probe overnight, at 38oC in humidified oven.
- Stringently wash slides in various concentrations of SSC.
- Proceed with detection of hybridized probe using Anti-DIG antibody. As per Roche method.
List all resources required including plant,
chemicals, personal protective clothing and
equipment, etc
 Lab gown, closed shoes, gloves;
 Cryotome in Histology lab (see slide prep method);
 De-waxing apparatus in Histology lab (see Histology unit);
 Glass slide; yellow “starfrost”;
 Cryotome blade;
 RNase free reagents; tips, pipettes and glass equipment’
 Coplin jars. Plastic “Blue lid” 50ml tubes.
 Glass coverslips,
 Wax Daco Pen.
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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
 When cutting slides possible to cut one-self. Follow procedures as outlined by Histology lab.
 Exposure to fixative, paraformadehyde can be harmful. Use in fume-hood in rm 207.
 Exposure to Formamide can be hazardous. Wear gloves, gown.
List emergency shutdown instructions
 In the event of inhalation or ingestion the first aid officer should be consulted immediately.

First Aid Officer: Renee Szokolai ext 58497
List clean up and waste disposal requirements
 Paraformadehyde fixative is to be collected into “toxic waste” bottles.
 Unwanted glass slides to be disposed into “yellow” contaminated glass bins.
 All other waste deposit in biological waste bin.
List legislation, standards and codes of practice
used in the development of the SWP
NSW OHS Act 2000
NSW OHS Regulation 2001
Code of Practice for the Labelling of Workplace Substances
AS/NZS 2243.2:2006. Safety in laboratories. Part 2: Chemical aspects
AS/NZS 2243.3: 2006 Safety in laboratories Part 3: Microbiological aspects and containment facilities
AS/NZS 1336:1997 Recommended Practices for Occupational Eye Protection
AS/NZS 2161.1:2000 Occupational Protective Gloves – Selection, Use and Maintenance
Supervisory approval, training, and review
Supervisor: Prof E Burcher
Signature:
Plant custodian:
Signature
List competency required – qualifications, certificates, licensing, training - e.g. course or instruction:
COMMON SENSE
SWP review date: 28-4-13
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