Diva Software CST Module

To access the CST module, the instrument must be on and connected. Choose Cytometer > CST. Wait for the CST module to load up and also for the cytometer to be reconnected (look at the bottom right corner of the screenfor a green light that says “Connected”). Follow the section for the action you are performing.

Note: Makes sure the instrument and lasers have enough time to warm up (approx. 30 minutes) before performing any of the below.

Prepare CST beads

  1. Thoroughly mix the CST beads by vortex.
  2. Add a big drop of beads to 300ul of PBS.
  3. Vortex to mix.

Check Performance (Daily)

  1. On the LSR-II instruments, select the Load Tube Manually checkbox.
  2. Verify that the Setup Beads lot ID selected matches your current lot of Cytometer Setup and Tracking beads. If not, have the administrator import the bead lot for you or use the menu to choose the appropriate lot.
  3. Place the tube of CST beads onto the Sample Input Probe of the cytometer and slide the sample arm underneath the tube..
  4. If your cytometer is an LSRII, set the flow rate to low (adjust the fine adjust knob to the middle) and put it in RUN mode.
  5. In the Setup Control window, click Run.
  6. Sometimes a bit of air needs to be purged from the cytometer. If the large beads do not form a tight circular population, press the abort button and unload the tube. Press the “Prime” button, wait a few seconds then press “Run”. Repeat the “Prime/Run” process a few times then run the CST beads again.
  7. Once the performance check is completed, a message appears and you can view the report in detail.
  8. Remove the CST beads from the cytometer.
  9. The cytometer performance results appear in the System Summary.
  10. If CST passed the check, you can move on.
  11. If CST failed the check, clean the instrument with 10% bleach and di-water for 3 minutes each and repeat CST. If it fails the check again, contact the administrator.

NOTE: A common reason the CST fails is running the beads too fast (> XXXX events per second), not allowing the laser enough time to warm up properly (machine should be on approximately 30 minutes before performing CST) and lastly bubbles/dirty flow cell.

Updated: 12-07-07