Live Cell Tracking

Important

Take images in as few fields of few as possible, with as few conditions as possible. Live tracking generates large quantities of data.

  1. At least 1 hour before imaging, turn on CO2 tank. Turn top knob to be completely open, and open valve so that it is inline with the tubing. Left gauge should read ~20 psi, right should read over 600 psi.

  2. Follow directions for startup.

  3. Ensure water tank is full to max line. Refill with sterile water and a serological.

  4. Replace stage with incubator stage (completely enclosed) if necessary.

  5. Turn on stage sensing (often the empty plates by the computer are better for sensing).

  6. Use the experiment suitable to your fluorescent needs (under the Fluorescence tab on the right panel).

  7. Find PFS offset (under PFS tab in rightmost panel).

  8. Click on Job Wizard (on the left??).

  9. If using an experiment from a previous run, right click the experiment and click continue editing. If you click run and edited anything, you must save it under a new name.

  10. Turn on autofocus on brightfield (or whatever channel is constant between all conditions).

  11. Autofocus on one point.

  12. Turn on use autofocus when PFS fails.

  13. Do a test run.

  14. Let the plate sit for ~20 min before starting image collection.

  15. To do media changes, hit the pause button in the corner (hit resume when plate is back in place).

  16. When live track is completed, follow directions for shutdown.