BD LSR Fortessa Standard Operation Protocol --- Turn on and turn ...

<strong>BD</strong> <strong>LSR</strong> <strong>Fortessa</strong> <strong>Standard</strong> <strong>Operation</strong> <strong>Protocol</strong>--- <strong>Turn</strong> on and turn off system (experienced user only)<strong>Turn</strong> on system:• Refill sheath tank and empty waste tank if necessary.• Please make sure all tubing are connected properly on sheath tank and wastetank.• <strong>Turn</strong> on main switch of <strong>Fortessa</strong> (wait for at least 20min before CST)• Stabilize fluidics: load 3ml of H2O then set “RUN” and “LO”.• <strong>Turn</strong> on computer and log in with Administrator• Unplug the internet cable (blue color)• Double click Cytometer Status and <strong>BD</strong> FACS Diva Software icons on desktop• Remove air bubbles from the sheath filter.• Log into FACS Diva software with your own login name and password.• The system is ready to use when it shows “Cytometer connected”• Please refer to SOP of CST if CST is needed.• Reconnect the internet cable for data transfer.<strong>Turn</strong> off system:• Make sure the proper cleaning procedure has been completed.• Log out Diva software.• Quite Diva software and close cytometer status window.• <strong>Turn</strong> off computer.• <strong>Turn</strong> off main switch of <strong>Fortessa</strong>.• Refill sheath tank.• Empty waste tank.

<strong>BD</strong> <strong>LSR</strong> <strong>Fortessa</strong> <strong>Standard</strong> <strong>Operation</strong> <strong>Protocol</strong>--- CST (experienced user only)• Click “Cytometer” > “CST” in Diva Software• Prepare CST beads (350ul FACS flow + 1 drop of CST beads)Note: please vortex CST beads before use and put CST beads bottleback to Fridge immediately after use.• Label CST beads tube with date and “CST” and Lot Number.• Press “RUN” and “LO” on Fluid control panel.• Check the beads lot number in Diva to make sure it is the same withthe beads you prepared.• Click “run” in Diva software.• CST will take about 7-10min. Please do not let CST beads run out.• Unload CST beads tube when finish and reload H2O tube.• Click “File” > “Exit”.If CST is not passed, please try the following steps and try CST again:1. Prepare one tube of 2ml FACS Rinse2. Prime X3 with FACS Rinse3. Run 10 min at “HI” flow rate4. Prime X1 with DI H2O5. Run CST again

<strong>BD</strong> <strong>LSR</strong> <strong>Fortessa</strong> <strong>Standard</strong> <strong>Operation</strong> <strong>Protocol</strong>--- Empty Waste Tank• Verify that the flow cytometer is in standby mode. Press theSTNDBY button on the control panel if necessary.• Click “Cytometer” > “standby” in Diva Software.• Disconnect the orange waste tubing and the black level sensorline from the waste container.• Take the waste tank to the sink.• Remove the lid of waste tank.• Empty the waste container.• Add approximately 0.5-1L of bleach to the waste container andtighten the lid of it.• Reconnect the orange waste tubing and level sensor line.• Reconnect Diva software with workstation by click “Cytometer”> “Connect”

<strong>BD</strong> <strong>LSR</strong> <strong>Fortessa</strong> <strong>Standard</strong> <strong>Operation</strong> <strong>Protocol</strong>--- Refill the sheath container• Verify that the flow cytometer is in standby mode. Press the STNDBYbutton on the control panel if necessary.• Click “Cytometer” > “standby” in Diva Software• Disconnect the air line (green) and fluid line (Blue) from the sheathcontainer.• Depressurize the sheath container by pulling up on the vent valve.• Remove the sheath container lid. Unscrew the clamp knob and pushdown to loosen, if necessary. Tilt the cap to the side to remove it fromthe tank.• Add sheath fluid to the sheath container (please use the blue bottle).• Replace the sheath container lid. Close the sheath lid and tighten theclamp knob to finger-tight.• Reconnect the air line (green) and fluid line (Blue).Please pay attention to fluid line (Blue) to make sure it is not twisted orstretched.• Reconnect Diva software with workstation by click “Cytometer” >“Connect”.• Remove air bubbles from the sheath filter.