CELLQUEST SETUP FOR CELL CYCLE WITH PI

1. Double click on your lab’s TEMPLATES folder.
2. Select your experiment template for cell cycle.
3. Go to ACQUIRE in the tool bar and scroll down, click on these 2 items:
        B. COUNTERS
4. Go to CYTOMETER on the menu bar and scroll down click on these 3 items:
        A. DETECTORS AND AMPS (Set all to “Lin” Mode)
        B. COMPENSATION (Set all to 0 and close)
        C. INSTRUMENT SETTINGS: If you already have Instrument Settings: in the file manager,
            select OPEN then look for your instrument settings by clicking DESKTOP then finding your
            lab’s INSTRUMENT SETTINGS folder and scroll down to find appropriate settings (or a
            previous data file), click OPEN, then SET, then DONE. This will load the settings into the
5. In the BROWSER window tell the computer where to store and what to name your data files.
        A. Click on the top CHANGE button, then in the file manager select DESKTOP, then your lab
            DATA folder, choose or create a folder where you want to save your data then click
        B. Click on bottom CHANGE button, then in the FILE NAME PREFIX pull down menu, select
            “SAMPLE ID”.
                 i. In the FILE COUNT box enter number “1” and click OK.
        C. In the Sample ID box, type in a name for your samples. This will be your file name so be
6. Go to ACQUIRE in the menu bar and scroll to ACQUISITION AND STORAGE
        A. Enter the number of events you want to save in the COLLECTION CRITERIA box (e.g.
            10,000 of R1).
7. Place the control on the cytometer and put in run. Click “ACQUIRE.”
        A. Using the FSC Amp and SSC Voltage and Amps, adjust the FSC and SSC so your cells are
            properly centered in the bivariate plot.
        B. Using the FL2 and FL2 W Voltage, adjust the negative cells so they appear at 200 on the
            linear scale.
8. Once you have your instrument settings set, click “pause,” “abort,” uncheck “setup” then click
    “acquire” to store a data file.
9. Vortex sample, place cytometer in RUN, mount tube on the cytometer and click ACQUIRE.
10. When the computer beeps and the counters stop, the data file is automatically saved. Proceed to
    change the SAMPLE ID and repeat step 9.
11. To save the data file before it has reached the total event count click PAUSE then SAVE in the
    Acquisition control window.

   Example of a Cell Cycle Template:

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