The Olympus FV1200 Z Drift Compensation (ZDC) module is an auto focus tool that corrects for changes in the distance between the objective lens and the specimen to compensate for focus drift during long time-lapses. It is available in two operating modes, single shot Single Shot (aka. focus searchFocus Search) mode or continuous focus Continuous Focus mode. Only Single Shot Mode is compatible with Z series.

ZDC in Single Shot Mode

In Single Shot (aka. Focus Search) Mode the ZDC activates briefly before image acquisition.

1. Start the microscope as usual but make sure the confocal software is closed
2. Focus on your specimen down the eyepieces

3. Enter the System Setting screen by pressing the spanner icon on the I3-TPC touch pad controller

4. Press the Customized tab on the System Setting screen
5. Make sure the Focus Limit Setting box is ticked and press the Detailed Setting button
6. Look down the eyepiece and focus slightly above your sample; beyond where you want any Z series to end
7. Press the 'All the same values SET' button to set the focus limit for all objective lenses
8. Press the OK button and then press the X button in the top right to close the System Setting dialogue box
9. Push the ZDC dichroic mirror (DM) lever on the left of the stage into place (see image below)
   
10. Open the confocal software and adjust the laser, detector and scan parameters until they are suitable for your specimen
11. Press the ZDC Setting button in the Acquisition Setting control panel
    
12. In the ZDC Control window, make sure the Search Zone is set to 100 µm and the 'ZDC control executes immediately before scanning start' button is ticked
13. Scan a reference plane in your specimen and press the Set Offset button in the ZDC Control panel (red arrow in image A below). The ZDC will search for the auto focus (AF) plane and will beep once when it finds it. The Offset is the distance from the AF plane to your reference plane
    
    
14. The Single Shot AF buttons are activated once the the Offset has been found (image B). These allow you to move between your current plane, the offset position (which is your reference plane) and the AF plane, which is the reflective surface of the specimen vessel
15. For Z series, set up Start and End positions and StepSize
16. For TimeScan time series, set up the Interval and number of timepoints
17. To operate ZDC during TimeScan, tick the 'Enable ZDC AF during Time Series Scan' box in the Acquisition Setting control panel
    
18. Press run experiment to start your time series (AF status will show “Searching…” during Focus Search)
19. When the acquisition has finished click series done

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