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LSR Operation Quick Reference

Note: Laser warm-up times: Argon (blue): 5 minutes. Diode 375 nm (Near UV): 5 minutes. HeNe (red): 15 minutes.
Lasers do not reach stable maximum output until they are fully warmed up. Although you are able to run samples after a minute or so, to ensure best stability, you should turn on the instrument slightly ahead of time to allow the laser(s) to warm up.

Startup.

  • Ensure that you signed up for instrument use using the on-line calendar (even if it is night or weekend).
  • Before switching on (or when starting your run):
    • Check sheath reservoir. A full tank provides about 10 hours' run time. Refill as necessary from large plastic canister near sink.
    • Check waste reservoir. Empty if it seems full. Discard contents into sink, flush with water. Put approx. 100 ml iodine based disinfectant (Mikroklene) into empty waste tank before returning it to the machine. ☠ DANGER ☠ Do not use bleach!
  • Switch on instrument, if not already on.
  • Switch on computer. If the computer produces a message saying that it cannot connect to the cytometer, restart the computer.
  • Turn on UV laser if desired.
  • Take off tube from SIP. Discard tube.
  • Press PRIME button and wait until instrument goes to STANDBY.
  • Turn sample fine adjust knob fully counterclockwise then turn clockwise 5 turns (this centers the control)
  • Make a new tube containing 1 ml (1/2 inch/ 1 cm depth) filtered DI water and put onto SIP.

Run.

  • Launch desired acquisition document. (Apple menu→Acquisition setups).
  • Double-click CytConnect icon. This runs a macro that connects to the cytometer, opens windows you will need and loads a set of default instrument settings. The macro gets confused if you move the mouse or type anything while it is running; if you do this you may have to restart the computer, so be patient until the instrument settings have loaded.
  • Create and select a folder in which to store your data.
  • Create a filename or prefix. Set file count to 1.
  • If you have run the same type of experiment before, use the Cytometer menu to load instrument settings from a previous data file. Don't forget to click SET!
  • Ensure the Setup box is checked.

Shutdown/finished.

  • Set fluidics switch to HIGH.
  • Potentially biohazardous samples: Run TriGene (1:50 or 1:100) for 5 minutes followed by three changes of DI water for 1 minute each.
  • Sytox: Run 100% bleach or 70% ethanol for 5 minutes followed by three changes of DI water for 1 minute each.
  • Make a new tube containing no more than 1 ml (1/2 inch/ 1 cm depth) filtered DI water and put onto SIP.
  • Run for 2 minutes.
  • Transfer your data to the CCMI data server during the clean-up procedure.
  • Set fluidics control to STBY
  • If you are the last user of the day:
    • TURN OFF LSR unit and computer.
    • Do not allow LSR to run overnight! Charge for instrument left on overnight is 50% of the time until it is turned off