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BD LSRIIs and BD LSR-Fortessa Maintenance Guidelines

  1. Login into Windows using your RUNet Login and password
  2. Check if the on-screen user interface shows BD LSRII (or BD LSR-Fortessa) and FACSFlow system “ON”. If it is “OFF” be sure to click only upper button once to switch “ON” both BD LSRII (or BD LSR-Fortessa) and FACSFlow system
  3. Please make sure that the green lights on both – instrument BD LSRII (or BD LSR-Fortessa) and FACSFlow system are actually “ON”
  4. If the fluorochromes / dyes you use in the experiment require the UV or 445 nm lasers to be “on” please make sure to follow the appropriate steps:
  5. If you need to change filter(s) on particular channel(s), for the first time please make sure to inform FCRC staff in advance and get proper training
  6. Before you start running samples please:
    • Check if the BD LSRII fluidic system is functioning properly:
      • Remove the tube with MQ water from the sip (Sample Injection Port)
      • Push “RUN” and “High” fluidic control buttons
      • Check if the buffer starts dripping from the sip:
        • If “yes” – please proceed to the next step
        • If “not” – please push the “Standby” button immediately and report the problem to FCRC Staff (see the notes on the end of the document)
    • Check if the trap filter attached to the pressurized plastic tank is free of air bubbles by bleeding sheath fluid from two ports:
      • Open the roller clamp on the top of the trap / bubble filter for 3-5 seconds
      • Sequentially open the second roller clamp (at the right side of the instrument) for 3-5 seconds
      • Check if you are getting consistent stream release from both ports:
        • If “yes” – please proceed to the next step
        • If “not” – you should suspect pressure problems or presence of bubbles, please immediately push the “Standby” button and report the problem to FCRC Staff (see the notes on the end of the document)
    • Wash the instrumentbefore you run samples:
      • “Run” at “High” 2 ml of 10% Bleach for 5 minutes
      • “Run” at “High” 2 ml of MQ Water for 5 minutes
      • Push “Standby” button and leave tube with 1 ml of MQ Water on the sip
        Note: Please be sure in all the cases not to exceed the maximum allowed volume of 2 ml in the sample tube
  7. Login into DiVa. Always choose to use CS&T settings. Create new experiment, or import the old one and duplicate without data. Run your experiment. Export data. Copy them to the CFS. Delete your experiment from the DiVa Database
  8. When you are done with the experiment if you have used the UV or 445 nm laser be sure to switch it off. Refer to the step 7 of “The Operation Procedure for UV Laser on BD LSRII-1” or step 3 of “The Operation Procedure for 445 nm Laser on BD LSRII-2”, respectively
  9. If you change filter(s) on particular channel(s) for your experiment, please make sure to switch back to default settings and put the spare filter(s) back to the storage place
  10. When you are done with the experiment be sure to wash BD LSRII (or BD LSR-Fortessa) properly:
    • “Run” at “High” 2 ml of 10% Bleach for 10 minutes
    • “Run” at “High” 2 ml of MQ Water for 10 minutes
    • Push “Standby” button and leave tube with 1 ml of MQ Water on the sip
  11. Before you leave FCRC please be sure to switch instrument off if required. Policy of keeping instruments “ON” or “OFF” depends on the time of the day:
    • During the daytime, we keep the analyzers “ON” all the time
    • If you finish your BD LSRII experiment after 6 pm on the weekday or any time on the weekend and if the next person is not physically present at the site we request to switch “OFF” the BD LSRII and FACSFlow system by using the on-screen user interface. Please click only upper button once to switch “OFF” both BD LSRII and FACSFlow system
  12. Please be sure to logoff Windows before you leave FCRC to avoid unnecessary charges. And it also prevents other researcher from usage of the instrument, since in 10 minutes Windows will automatically lock the session under your name

 

Note: Please report all the problems/concerns to FCRC Staff:

During business hours — go to DWB205 and tell the FCRC staff member
After hours — e-mail to FCRC_Staff@rockefeller.edu; leave the note about the problem on the instrument’s keyboard; proceed on step 10 from the above list