CytoFLEX Start-Up and Q/C

Start-Up

1. Turn on the power switch on the back cover of the instrument, located just above the power cable.

2. Log on to the computer and double-click to start CytExpert.
a. Ensure that the Connected icon on the Status Bar near the bottom-left side of the display is green.
b. If the icon is not green, ensure that the instrument USB is securely connected to the computer and restart the computer.
3. Select System Startup Procedure in the Cytometer menu and follow the software prompts.


Instrument QC

1. Perform Daily QC:

  • Select the QC/Standardization Menu which will load the QC module
  • Here select QC under the Task and from the drop down menu on the Lot No. select the appropriate lot number for the CytoFLEX QC beads
  • To prepare the QC beads:
    0.9ml water + 0.1ml FlowClean Agent + 3 drops of CytoFLEX Daily QC beads then vortex. Diluted beads can be stored at 2-8 degrees C in dark for 5 days.
  • Initialize the system if it has gone into Standby mode then select Run. QC wizard will begin to run the QC Report and it will pass or fail once the program completes.
  • If FAIL, repeat QC after verifying: are beads expired?, was sample prepared accurately?, prime flow cell, follow Daily Clean and cleaning Procedures then retest. If FAIL occurs after troubleshooting and running QC three times in a row contact BC Customer hotline.
  • QC WIZARD CHECKS LASER POWER, LASER DELAY TIMES, and APD performance.
  • After completion close out the QC module by selecting ‘Close QC/Standardization’ under File menu.

 

Importing Target Library for QC

  • While in the QC/Standardization mode select ‘Target Library’ under the ‘Settings’ Menu.
  • Download target library if internet connection on cytometer.
  • If not, go to Target Values.
  • Then choose these categories: Research & Discovery-- Flow cytometry—Instruments—CytoFLEX--CytoFLEX QC Fluorospheres Target—All—English—Search--CytoFLEX QC Fluorospheres target values—
  • Download and Save to the desired file path.

QUICK TIP For each new Lot number, download the corresponding target value file and refer to Step 2.

  1. Using the Lot No. drop-down list on the left side of the screen, select the Lot No. corresponding to the QC fluorospheres.
  2. Select , insert the prepared QC sample tube into the sample tube holder, then select to start QC.
    NOTE QC may take several minutes. The minimum flow rate for the QC fluorospheres is 100 events/ second. The results are displayed and saved automatically once it is completed.
    • Green icons in the Result section indicate Pass.
    • Red icons in the Result section indicate Fail.
  3. If QC fails, prime the instrument, initialize, and repeat QC.
    NOTE If QC fails two times in a row on the same day, contact your Beckman Coulter Representative.

Creating Levy-Jennings Charts

  • Under the QC/Standardization module select the ‘LJ icon’ from the left edge of the screen (the icon resembles a graph). LJ charts settings will open.
  • In the Laser Tab, select the power and/or delay checkboxes for each laser as needed
  • In the channels tab, select each channel checkbox as needed.
  • Click Apply, then OK. Select the plot, choose start and end dates. 
  • Close the QC module under the ‘File’ menu