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1. Load 10% bleach. Change flow rate to 11 and run for 5 min.
2. Repeat with 5% Decon 90.
3. Repeat with DI water.
4. Stop stream.
5. If the next sort booked on the instrument in that week uses the same nozzle
   1. Load a full tube of 70% EtOH and initiate clean flow cell command. 
6. If the next sort booked on the instrument in that week uses a different nozzle
   1. Remove current nozzle. 
   2. Replace with cleaning nozzle.
   3. Load a full tube of 70% EtOH and initiate clean flow cell command. 
   4. When complete - remove cleaning nozzle and remove nozzle clip.
7. If there is no other sort booked for the week. 
   1. Remove current nozzle. 
   2. Replace with cleaning nozzle.
   3. Initiate fluidics shutdown program from cytometer menu.
   4. Complete fluidics shutdown. 
   5. Remove cleaning nozzle and nozzle clip and replace to default locations.
8. Spray the sort chamber and other appropriate internal chambers specific to the instrument as well as exposed appropriate external surfaces with 70% w/v ethanol and wipe clean. Spray again with 70% w/v ethanol and allow a contact time of at least 10 minutes.
9. Wipe the areas and follow with spraying and wiping clean with 70% ethanol
10. System is now ready to be shut down. Turn off instrument, gas, chiller, AMO & BSC afterwards. Log off computer.
11. Relieve any pressure from the tanks by venting using the venting valve.

Data management

• Under no circumstances should there be any patient/person identiable data recorded on the instruments as the data is not privately held and is accessible to all researchers. 
• User data is deleted after 7 days off the instrument computers, it is the responsibility of the user to have transferred data to a secure location.
• WRHFlow recommends exporting data from BD FacsDiva as FCS3.0 files in linear format to D:\BDExport\FCS. This folder is automatically synced to the WIMR Scientific Platforms Drive that all WIMR active directory users can access. For the sync to work properly your experiment must be in a folder with "FirstnameLastinitial" syntax. Please note this folder is accessible to all researchers.
• If you would like your data accessible from outside of WIMR, a user specific share can be setup. This works by creating a copy of a specified folder on university provisioned storage that is shared with the user. This requires an email to wrhflow@sydney.edu.au and it is the users responsibility that all ethics approvals are complied with. 

Application Settings

Application settings are useful in ensuring longitudinal studies (studies measured over multiple days/weeks/months) maintain consistent MFIs over the course of the experiment. We have successfully implemented and assisted many users in creating and applying application settings to their experiments.

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1. If you are in doubt of your ability to clean a chemical spill safely then evacuate the area and seek help. It is better to ask for help if you are not sure of how to clean it up properly.
2. If there is risk to the rest of the building contact the emergency contacts to initiate building evacuation.
3. Review the SDS and make sure you understand the hazardous properties of the spilled material before you attempt to clean it up.
4. First aid is always the top priority. If a hazardous material is spilt on yourself, remove any potentially contaminated clothing immediately and use the emergency shower. If a chemical spill has entered your eyes flush for at least 15 minutes at the eyewash station. Seek appropriate medical treatment.
5. Determine whether it is a major or minor spill. A major spill is one that involves a large amount of hazardous material, poses a risk of fire/explosion, a respiratory hazard exists, or when dealing with unknown chemical spills. Seek assistance from emergency contacts for major spills. 
6. For minor spills alert lab manager, safety officer and others in the lab and cordon off the affected area. Isolate the spill.
7. Retrieve the spill kit (there are also adequate instructions in the chemical spill kit). Located in the shared prep lab J2.06 - see appendix for location. Stop and think about your plan to clean the spill. Remove the gloves and goggles and from the kit, put them and all appropriate PPE on before approaching the spill.
8. Identify the spill - there is a pH strip tester that can be used, also can refer to SDS, labels, supervisors. 
9. Select the agent to clean up the spill.
   1. For acid spills: Spill-X-A® acid neutraliser
   2. For caustic spills: Spill-X-C ® caustic neutraliser
   3. For solvent spills: Spill-X-S ® solvent adsorbent
   4. For formaldehyde spills: Spill-X-FP ® formaldehyde
      polymeriser
   5. In some instances combinations of adsorbents many be necessary. 
10. Encircle spill, cover with adsorbent
11. Mix adsorbent into spill
12. Using the absorbent pads from the spill kit, carefully wipe up the
    spilled liquid, again working from the outside in.
13. Beware of any neutralising reactions
14. Collect powdery residue using spatula, deposit in waste bags
15. Place all waste materials in a plastic bag. Once the spill has been
    fully cleaned, place the waste bag with in the fume hood temporarily.
16. Label and dispose of waste according to building policies. 
17. Remove PPE and thoroughly wash hands.
18. Report the spill using the buildings incident notification form.

Emergency shutdown for stream failure:

If a stream failure occurs, the AriaIII may detect this and shut off the stream and sample automatically. If this occurs during sample being put through the instrument, the instrument must be given time for any potential aerosols to settle or be removed via the biological safety cabinet before accessing the sort block.

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1. Spray the sort chamber, sort collection block and other appropriate internal chambers specific to the instrument with 1:50 100 TRIGENE and wipe clean. Spray again with 1:50 100 TRIGENE and allow a contact time of at least 10 minutes.

1. Determine cause of sort failure if possible.
2. Remove nozzle and sonicate if necessary.
3. Restart the stream if possible.
4. If nozzle clog occurred – refilter sample or use larger nozzle before loading sample.
5. Log the failure in the sort log.Emergency shutdown if an electrical hazard is present

If there is an electrical hazard and the instrument is plugged into a power point that is not on the uninterruptible power supply (blue power point), pressing the emergency power off button will stop electricity to the instrument. Note if an electrical device is plugged into the blue power point it will continue to be live. In this event leave the room, ensure a message is placed on the door alerting others and seek assistance.

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1. .

Associated Documents

This Standard Operating Procedure should be read in conjunction with:

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