Usage Policy: Cytometry

1. Startup and shutdown of the machines is performed by Imaging & Optics Facility staff only: we cannot make exceptions, as they result in down-times and additional costs.
2. FACS Aria III service can only be provided within the following times: 9 am till 5:00 pm Monday-Thursday, 9 am till 15:30 pm Friday.
3. To ensure availability of the FACS service, your reservation needs to be made at least two working days in advance of your experiment (i.e., Monday for Wednesday etc.). Only when the Aria is booked on time, can we guarantee that the service will be provided – personnel has to be scheduled for the startup/reconfiguration/shutdown times. (Weekend days count as one day- i.e. , you have to book on Friday to ensure service on Monday.)
4. If FACS service is requested on a shorter term (email: cytometry[at]ist.ac.at), we TRY to match your needs, but cannot guarantee it. The reason is, that our FACS experts are also providing other services and have to plan their time in advance.
5. Cancelling FACS bookings within 24 hours prior to slot start is only possible via email: cytometry[at]ist.ac.at
6. Since the device has to be prepared well in advance, failure to cancel at least 2 hours before the start of your session will result in startup and shutdown charges.
7. In case late sorts (past 6 p.m.) or weekend sorts are required: we cannot guarantee them. In case there is a SERIOUS experimental demand on this, we can discuss it on a case-by-case basis and look for a solution. Please contact us about such requests several weeks in advance.
8. If your configuration (nozzle size, temperature or sample collection rack) differs from that of the previous user or if you are the first user at the time of booking, please make sure you always select the ‘Set sorting configurations by IOF’ checkbox in your booking. This is needed to ensure enough time before your slot to allow us to reconfiguration the sorter for your experiment– otherwise we’ll have to do it in your session!
9. Presence at machine: Due to the complexity of the system, Users are required to be present while sample is being measured at all times.
   

Cytometry service hours for assistance and for novice users

• Monday – Thursday: 8:30 – 17:30
• Friday: 8:30 – 15:30

Usage policy

• Novice: All users start with ‘novice usage rights’: Bookings can only be made during the above mentioned cytometry service hours, and need to be confirmed (see guidelines) → Startup, shutdown and sample line exchange must be completed within this time-frame.
• Autonomous: to book devices outside of core hours “autonomous usage rights” are required. To obtain autonomous user rights on the Sony SH800, the handling is evaluated over the 5 initial independent sessions that need to be done without need of experimental assistance (but for technical troubleshooting) → incidents due to incorrect use, startup, or shut down procedure might result in a prolongation of the novice period.
• Contaminating samples: all experiments with potentially contaminating samples (e.g. bacteria) MUST be done with a separate (user owned) sample line. Own sample lines can be used when desired.
• Sample line exchange: by default, the sample line exchange needs to be requested (see above) and is performed by IOF staff (during core facility hours). Autonomous users can request an additional training (see communication channels) that allows sample line exchange by experienced users.
• Presence at machine: Users are not required to be present at the machine for the entirety of their booked session. However, they are responsible for proper usage and are expected to regularly check in on it.
  If a problem is encountered that might be damaging to the machine (e.g. leakage or clogging) and the user is not available, the run will be stopped and session ends.
• Sample running out: The System does not have a bubble detector! Always avoid sample from running out (air gets sucked into system).
  If air was sucked into the machine, multiple sample line flushes and chip de-bubbles should be performed first. If this does not resolve the issue, fluidics calibration and QC have to be repeated. While not required, we recommend using a new chip, as air might get stuck in the chip.

Assistance & Requests (help, sample line exchange, etc.)

• To ensure availability of FACS service (novice assistance), your reservation needs to be made minimal two working days in advance of your experiment (i.e. Monday for Wednesday etc.). Only when service is booked in timely in advance, the Imaging & Optics Facility can guarantees that service will be provided. (Weekend days count as one day – i.e. you have to book on Friday to ensure service on Monday.). Make sure to book assistance using the correct form in PPMS: Request assistance by IOF staff for Flow Cytometry
• Assistance/session is guaranteed upon confirmation of your request or booking (PPMS – novices).

End of a session / shutdown Sony SH800SFP

• Cleaning and shutdown protocol must be followed strictly and completely → Tab 4 in the manual
• Clean the sample chamber and sorting chamber with 70% EtOH → ensure that any sample remains and salt crystals are removed, especially from the deflection plates
• Refill DI water reservoir after every session
• Empty waste tank after every session
• Shut down the main device, PC and compressor completely, unless you are directly approached by the next user → Tab 4 in the manual
• The last user of the day must refill the sheath tank with PBS to the level indicated on the tank (yellow label) → add the amount of used PBS and MilliQ water bottles to the consumables log-book (located in the pouch inside the cover of manual)

Cytometry service hours for assistance and for novice users

• Monday – Thursday: 8:30 – 17:30
• Friday: 8:30 – 15:30

Usage policy

• Novice: All users start with ‘novice usage rights’: Bookings can only be made during the above mentioned cytometry service hours, and need to be confirmed (see guidelines) → Startup and shutdown must be completed within this time-frame.
• Autonomous: to book devices outside of core hours “autonomous usage rights” are required. To obtain autonomous user rights on the CytoFLEX LX, one needs to complete 5 independent sessions without need of experimental assistance → incidents due to incorrect use, startup, or shut down procedure might result in a prolongation of the novice period.
• Contaminating samples: all experiments with potentially contaminating samples (e.g. bacteria) MUST use the dedicated shutdown procedure in the provided manual.
• Presence at machine: Users are not required to be present at the machine for the entirety of their booked session. However, they are responsible for proper usage and are expected to regularly check in on it.
  If a problem is encountered that might be damaging to the machine (e.g. leakage or clogging) and the user is not available, the run will be stopped and session ends.
• Sample running out: The System does not have a bubble detector! Always avoid sample from running out (air gets sucked into system).
  Measured sample volume in stopping rules should be set as 40µl less than actual volume to account for dead-volume (Plate mode).
  If air was sucked into the machine, it is mandatory to re-run system start-up program before continuing (~10minutes). QC does not need to be repeated.

Assistance & Requests

• To ensure availability of flow cytometry service (novice assistance), your reservation needs to be made minimal two working days in advance of your experiment (i.e., Monday for Wednesday etc.). Only when service is booked in timely in advance, the Imaging & Optics Facility can guarantees that service will be provided. (Weekend days count as one day – i.e. you have to book on Friday to ensure service on Monday). Make sure to book assistance using the correct form in PPMS: Request assistance by IOF staff for Flow Cytometry
• Assistance/session is guaranteed upon confirmation of your request or booking (PPMS – novices).

End of a session / shutdown CytoFLEX LX

• Cleaning and shutdown protocol must be followed strictly and completely → Tab 4 in the manual
• Replace sheath Tank and refill used one with CytoFLEX sheath fluid.
• Empty waste tank after every session