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| Task or scenario | Hazard/s | Associated harm | Existing risk controls | Current risk rating | Additional risk controls? | Residual risk rating? |
|---|---|---|---|---|---|---|
| Instrument operation | Electrocution | Contact with electricity can cause electric shock and burns | Routine instrument maintenance - to ensure instrument is in good condition and cabling is not damaged. Electrical equipment annual testing. Educate users to check for visible liquid leaks. Safety circuit breakers and fuses on instrument to prevent general electrocution due to instrument failure, especially in the presence of liquid. Emergency power off button located in laboratories to disconnects power to the red power points and not the blue uninterruptible power points. Routine maintenance - to ensure instrument is in good condition Bright LED - lit to notify of high voltage deflection plates - educate users about importance of ensuring high voltage is off before accessing plates and surrounding area Instrument covers - when sorting all covers are to be in place as physical access is restricted High resistance installed on HV plates - to limit current draw Circuit breakers and fuses on instrument - to prevent general electrocution due to instrument failure, especially in the presence of liquid | low | ||
| Contact/exposure with biohazardous materials | Exposure to biohazardous material can cause health issues | Electrostatic droplet cell sorter creates thousands of drops per second. Droplets can be small and readily enter airways. Therefore containment of aerosols is critical. This is achieved via having the cell sorter in a BSC, operating with covers, ensuring instrument aerosol management option is properly utilised and following emergency procedures in the case of sort stream failure. BSC - Instrument is inside a biosafety safety cabinet for increased protection. Software - Software controls to maintain a stable stream Filtration - Ensure samples are filtered prior to loading on the instrument, avoid blockage to minimise aerosol generation during sorting Visual check - Check sample for visible clumps that can cause nozzle clogs Signage - Emergency sort failure procedure in SWP and in room Signage - Signage on door during sorting to prevent unauthorised access during sort. Aerosol management option installed and in use as per SWP. PPE while emptying waste tank and adding bleach to waste tank (bleach decontamination of waste material). Engineering control - SIP sheath cover installed to minimise aerosol exposure from sample upon unexpected sample unload. Ensure users and support technicians are familiar with risk assessment and SWP for the material used. PPE – gloves, gown & enclosed shoes (P2 mask and safety glasses in sort failure). Users empty waste upon setting up of instrument with running water gently down the sink. 100ml of bleach is added to the instrument waste container after emptying the waste. Biological spill kit - Access to emergency biological spill kit and/or cleaning equipment. Bleach / decon / ethanol decontamination of sample lines. Project approval process. Handling samples (e.g. transferring, pipetting) in biological safety cabinet. Running the sample dry can introduce air bubbles resulting in unstable stream and rogue aerosol formation. SWP states to not run the sample dry. | low/medium | air bubble detector in sample line to prevent running sample dry. | low | |
Manual handling (i.e. lifting, transferring) heavy weight such as waste tank | Manual handling can result in injuries of the back, neck, shoulders, arms or other body parts | Providing information and training to workers on manual handling tasks and request for assistance options Maximum possible weight for tanks is <10kg Trolley/pallet jack - to transfer more than 1 box of saline/water – lifting only 1 box at a time Education - Providing information and training to workers on manual handling tasks Planning - Organising manual handling tasks in a safe way, with loads split into smaller ones, and proper rest periods provided | low | |||
| Failure to adhere to SWP | Exposure to laser | High power lasers used in instruments can cause skin/eye damage/burns | Use laser safety shielding at all times - to prevent avoid laser exposure. Do not disengage automatic shutters – electronically or mechanically activated when certain covers are open. Educate users not to circumvent shutters and to avoid looking into any exposed lasers or reflections as laser light can be invisible. Laser safety shielding - to prevent avoid laser exposure. Laser safety shielding - to prevent laser exposure Automatic shutters – pressure driven or mechanical when cover is open | low | ||
| Operating AriaIII – pinch hazard | Manual handling – pinch hazard while sample loading | Injury to hand | Education – Providing information, demonstrating and observing proper instrument use while loading samples on the ARIAIII during training to avoid pinching. Including closing of cover before loading of sample occurs. Instrument hardware monitoring for failed tube load. Physical cover - Cover to prevent accidental contact with the stage during sample load up Emergency stop button – easily accessible button to stop instrument in an emergency situation | low | ||
| Manual handling – pinch hazard while unscrewing/screwing tank | Injury to hand | Education – Providing information, demonstrating and observing proper instrument use while loading samples on the ARIAIII during training to avoid pinching. Including removing lid completely before filling tank and removing air supply line while depressurising lid for removal. | medium | Removal of air line until lid is being removed. | low | |
Handling hazardous chemicals including Bleach, Decon/Contrad, Ethanol, CST beads (Sodium Azide) | Hazardous substance exposure | Eye exposure causing eye damage Contact with skin can cause irritation or burn | Safety goggles are provided for researchers and recommended for use in the lab. Emergency showers & eye wash stations available in shared J.2.06 laboratory. Chemical spill kit available in shared lab J.2.06. PPE – gloves, gown & enclosed shoes are necessary for working in the laboratory. SDS available to users to ensure awareness of relevant chemical hazards and emergency procedures. Required to read and understand before using chemicals. | low | ||
Sample handling | Contact with bio-hazardous material | Exposure to bio-hazardous material | Handle samples in biosafety cabinet PPE – gloves, gown & enclosed shoes are necessary for working in the laboratory. Access to emergency biological spill kit and materials to clean up spills. | low | ||
High pressure gas | Physical injury caused by disconnected tube supplying high pressure air | Tubing connectors may degrade resulting in escape of air | Auto shutting off connections if disocnnected. Emergency shut off valves - Education of user to the location and operation of the emergency shut off valve. Venting ports - Education of users to depressurise any tank/fluidics line before accessing or opening the sheath tank and other components. | low | ||
| Exposure to sodium azide while performing CST | Exposure to toxic chemical | Acute toxicity | Introduce relevant hazards with SDS. Ensure PPE, i.e. Gloves are worn while handling the sample. Sodium azide is used for a minimal time during the quality control procedure. Ensure only minimum quantity (1 drop in 300uL) made each time quality control solution is made. Dual barrier protection (gloves, tube) & lid. | low | usage of nitrile gloves if possible - added protection | low |
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- Turn on the CAS BSC hood, chiller, high-pressure gas and aerosol management system (set at 20% - for operational velocity evacuation).
- Turn on the main switch to the ARIAIII located on the left of the instrument.
- Log into Windows using provided credentials.
- Log into the PPMS account with provided credentials.
- BD FACSDiva should launch automatically as should Google Chrome (contains the sorting log).
- Log in to BD FACSDiva, the software will detect and connect to the instrument. If it does not do it automatically, click Instrument in the main menu and connect the instrument manually.
- If a CST mismatch dialog appears always click ‘Use CST Settings’
- Open the BCS cabinet.
- Open the front cover of the AriaIII and prob it open.
- Determine if the correct nozzle is in place in the flow cell.
- If it isn't the cleaning nozzle is in place perform a fluidics startup (as this indicates the fluidics was shutdown).
- If the incorrect nozzle is in place in the flow cell, sonicate the appropriate nozzle in DI water for 5 minutes in a 5mL FACS tube. Ensure sonicator has sufficient liquid in it to conduct cleaning power to the nozzle.
- If needed - unlock and remove the previous nozzle or waste nozzle from the flow cell. Place the waste nozzle into an empty slot on the nozzle storage block.
- Use a Kimwipe to remove any residue liquid on the sonicated nozzle to be installed if necessary.
- Clean any salt residues around the nozzle assembly area using ethanol.
- Insert sonicated nozzle with the O-ring facing upwards. Lock the nozzle.
- Since the nozzle was changed, also change nozzle size settings in the software by setting the correct cytometer configuration - >Cytometer>Configurations>Select correct nozzle>Apply.
- Open the sort block door, click the stream start/stop button to turn on the stream.
- Check the stream video feed is not impeded by any liquid/debris on the lens. If this is the case it can be cleaned with a lens cleaning tip.
- Check the stream angle. If the stream flows away from the centre of the waste catch, loosen the screws on both sides of the collection assembly and rotate the sort block to adjust.
- Close the sort block and thumb screw it shut.
- If the nozzle wasn't changedcorrect nozzle is in place in the instrument, ensure the correct settings are instrument configuration is selected and applied in the software by looking at the window title bar.
- The ARIAIII flow cell cover should be open.
- Open the sort chamber (we need to view whether the stream is aligned or not.
- Close the front cover of the BSC.
- Be ready to change the waste catch position and then start the stream.
- Change the waste catch position if necessary.
- Stop the stream. This starts air into the sheath tank which is used to minimise contamination of the sheath tank when refilling.
- Unscrew the sheath tank lid completely.
- Remove the air pressurising line.
- Relieve all pressure from the tank by venting using the venting valve, and breaking the air-pressure seal by pushing down. Ensure you keep fingers clear of the lid and remove it. It can be stored temporarily on the fluidics cart (keeping the inner face not touching anything).
- Connect the air pressurising line promptly.
- Fill sheath tank to the weld mark (~10cm from top of tank).
- Reseal sheath tank - only holding the screw mechanism to keep fingers clear of the lid.
- Empty waste container slowly into the sink under running water. Put 100ml of bleach into the waste tank.
- Reconnect waste container.
- Check the filter attached to the sheath and at the front of the fluidic cart for air bubbles. If needed, ethanol spray and then bleed out any air bubbles if required from the finger screw vent on the filter.
- Start the stream again. The stream should be aligned in the centre of the waste catch.
- Close the sort chamber door.
- Open the front cover of the BSC hood and close the front of the ARIAIII lid.
- Close the front cover of the BSC.
- Inspect the stream for stable droplet generation, symmetrical droplets and similar breakoff position to the last time the stream was started. Click the sweet spot button and inspect the stream. Allow stream to stabilise. Adjust frequency and amplitude as needed with sweet spot off, updating values if necessary.
- Carefully check the instrument for wet areas indicating any leaks in the tubing or failing valves.
- Do not continue the sort with an unstable stream or leaks.
- Allow ARIAIII stream/lasers to warm / stabilise for at least 30 min.
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