OHS026
Safe Work Procedure
Faculty/Division
Medicine
Document number
SOMS GBD SWP 029
School/ Divisional Unit
School of Medical Sciences
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
ATP release in the Organ bath
Title:
Description: collect samples to measure ATP release from the tissue in organ bath
Associated risk assessment title and location: RA working with electrical equipments 017, RA waste management 015 located in
Room 209B, RA ATP release, RA using organ bath.
Describe the activity or process
Turn on computer and open POLYGRAPH program.
Turn on grass polygraphs.
Calibrate tensions of transducer by:
Click PARAMETERS, go to “Calibrate”, copy each “Volts” box to the felt bottom box - copy selected cell as voltage for
zero gram force to make the baseline calibration; then hanging up 20.5g weight to each transducer and copy each
”volts” to the right bottom box - copy selected cell as voltage for full-scale force to make the weight scale. Close the
calibrate window when these all be finished.
Make up Kreb’s solution and bubble the solution with carbogen for 10-30mins.
Pour solution to the top tanks and allow the solution to flow through each bath.
Turn on heating pump and allow the heated water system to flow through.
Dissect tissue strips under Helton hood and hang up the strips to connect the strips transducer.
Adjust the tension to 1.0 g by tightening the strings.
Make the tissue to rest for 1 hour and start the experiment.
Use chemicals, peptides or electrical stimulators to change the tension of the tissue and to record the tension changes by
POLYGRAPH.
Collect and aliquot samples of the organ bath fluid into eppendorfs stored on ice using a 1ml syringe. Freeze samples at -40°C until
time to measure ATP content.
Cut off the strips and thoroughly wash organ baths with water after the experiment finished.
Change, soak (in 70% ethanol) and wash the baths every two experiments.
Make the surface treatment of the baths by using Surfasil Siliconizing Fluid every two washes.
Clean up the working area and wipe out the contaminated area and instruments with 70% ethanol and then with water.
Dispose all gloves, strings, tips, leftover tissue to biological waste bin and autoclaved bin.
List all resources required including plant,
chemicals, personal protective clothing and
equipment, etc




Suitable PPE,
Chemicals, peptides
Computer and grass polygraph
Needle and shield, sharps bin
List potential hazards and risk controls including
specific precautions required



Tissue contaminated or infectious – wear suitable PPE, and always change gloves as soon as you have finished handling
tissue, use separate instruments that specially designated for animal tissue dissection.
Potential aerosol hazard – use dissecting cabinet when dissecting animal tissue, always wear suitable PPE.
Injured by dissecting instruments, needle for sample collection – All users are immunized against Hepatitis B, are trained
to use instruments, notify the first aid officer and go to see a doctor to get blood test immediately after injure.
___________________________________________________________________________________________________________
___________
Page 1 of 2
Safe Work Procedure
Date Effective: 01/01/2007
Uncontrolled document when printed
Current Version: 1.2, 15/08/2007
List emergency shutdown instructions

Computer and glass polygraph have “OFF” switch that can be used in emergency.

Shut all electrical equipment off at power point in case of emergency.
List clean up and waste disposal requirements
Not applicable
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 Labeling of Workplace Substances
Australian Standard AS2243.6-1990. Safety in laboratories. Part 6: Mechanical Aspects.
Australian Standard AS2243.7-1991. Safety in laboratories. Part 7: Electrical Aspects.
Safe Work Procedure Form (OHS026)
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:
SWP review date: 20-04-2013
Responsibility for SWP review: Fei Shang
___________________________________________________________________________________________________________
___________
Page 2 of 2
Safe Work Procedure
Date Effective: 01/01/2007
Uncontrolled document when printed
Current Version: 1.2, 15/08/2007