Mar 20, 2025
SOI - BD FACSAria Fusion Cell Sorter - Instrument Shutdown
This protocol is a draft, published without a DOI.
- 1European Molecular Biology Laboratory (EMBL)
Protocol Citation: Daniel Gimenes 2025. SOI - BD FACSAria Fusion Cell Sorter - Instrument Shutdown. protocols.io https://protocols.io/view/soi-bd-facsaria-fusion-cell-sorter-instrument-shut-d5x687re
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 19, 2025
Last Modified: March 20, 2025
Protocol Integer ID: 124638
Keywords: Flow Cytometry, Cell sorting, FACSAria Fusion, Core facility
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Abstract
This Standard Operation Instruction is intended to guide the facility users to perform a fluidics shutdown on a BD FACSAria Fusion‱ instrument
Guidelines
This protocol should be done by:
- The faciity staff
- Trained users of the FCCF.
This protocol should be performed:
- Everyday after the instrument is used.
Perform the steps subsequently unless otherwise stated.
Materials
- BD FACSDiva™ CS&T Research BeadsBecton Dickinson (BD)Catalog #655051
- BD FACS™ Accudrop BeadsBecton Dickinson (BD)Catalog #345249
- FACSCleanBecton Dickinson (BD)Catalog #340345
- BD FACSFlow™ Sheath FluidBecton Dickinson (BD)Catalog #342003
- FACSRinse (discontinued by the manufacturer)
- Distilled H2O
Safety warnings
Handle FACSClean carefully. It should't enter in contact with the skin and eyes (wear gloves, lab coat and glasses) and it ruins colorful clothes.
Before start
Be sure to wear gloves and lab coat at all times!
General rule: spray 70% ethanol in all connectors before plugging any tubing!
Exporting your data
Exporting your data
Export your FCS Files and PDFs to the FCCF data server.
If necessary, create a template for your experiment.
Delete your experiment from the "Experiments Browser".
Cleaning
Cleaning
Run sequentially Rinse, Clean and dH2O for 5 min in a flow rate of at least 4.0
Go to “Cytometer”, “Cleaning Modes”, “Sample Line Backflush”.
On the Sample Line Backflush window, press “Start”.
Let it run for 30 seconds, then stop the Sample Line Backflush by pressing “Cancel”.
Turning off the stream
Turning off the stream
Press the green checkmark button on the Stream Window to turn off the stream.
Performing Fluidics Shutdown
Performing Fluidics Shutdown
Go to “Cytometer” → “Fluidics Shutdown” Follow the instructions on screen.
Open the nozzle lock (by turning it anti-clockwise until 8:00 position), remove the nozzle and install the closed loop nozzle with the red o-ring facing up. Close the nozzle lock (by turning it clockwise until the 12:00 position) and confirm on the software by pressing “Done” for the first two options.
Rinse the nozzle orifice with dH2O: Pour dH2O (from the 50 mL syringe with a filter) into a 50 mL Falcon Tube. Grab the 5 mL syringe with the yellow tip and fill it with the tube water. Take the nozzle and turn it upside down (with the red o-ring facing down!). Place the tip over the nozzle orifice and gently rinse it until it’s possible to see liquid coming through in a form of a stream.
Dry the nozzle with compressed air or soft tissue paper (be careful to not displace the o-ring!!!). Store it in its corresponding plastic case.
Open the lower right metal door in the instrument. Pull the fluidics cart by pressing the small yellow lever on the right upper part of the cart.
Remove the fluidics tubing from the sheath filter and plug into the ethanol filter (red labeled connections). Remember to always spray 70% ethanol when exchanging the connections.
Note
Check the instrument folder for pictures if in doubt!
Remove the air connection from the sheath tank and plug into the steel ethanol tank (green labeled connections).
Note
Check the instrument folder for pictures if in doubt!
Keep the fluidics cart open. Click on “Done” for the next step. The Fluidics Shutdown will start and it take approximately 5 minutes.
Close the sort block
and the instrument coverings.
After the Fluidics Shutdown completes, vent the sheath tank by pulling up on the ring on the pressure release valve. Once done, press “OK” on the software.
Fill a 5 mL tube with 3 mL of dH2O. Put it on the loading port. Press “Done” to move to next step in the software.
Note
Keep the tube there after is done!
Confirm the Fluidics Shutdown procedure by clicking on “OK”.
Go to “Cytometer” → “Cleaning modes” → “Clean Flow Cell”.
With the tube containing dH2O still on the loading port, confirm by pressing “OK”.
If the 2.0 ND filter is installed in front of the FSC detector, swap it for the ND 1.5 filter.
Discard all cleaning tubes. If the Accudrops tube has leftovers, put it in the fridge for posterior use.
Open the waste tank and add 10 mL of Sekusept Plus.
Spray 70% ethanol over the lid of the sheath tank before opening. Unscrew the sheath tank cover knob and remove the lid. Refill it with sheath fluid from the 20 L FACSFlow cardboard box until it reaches the upper weld line inside of the tank. Put back the lid and tighten the knob.
Note
Do not overfill the sheath tank!
Push back the fluidics cart and close the metal door.
Turning off the instrument and cleaning up the workspace
Turning off the instrument and cleaning up the workspace
Close the DIVA software.
Turn off the computer.
Turn off the instrument by pressing the green button on the right side.
Close the window of the biosafety cabinet and switch it off by pushing the ventilation symbol button.
Turn off the cooling unit.
Organize the workspace: put all the tip boxes, pipettes, syringe, tube boxes, collection devices, etc in its proper designated spaces.