Molecular Devices ImageXpress Confocal HT.ai, User Manual

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Chapter 8: Maintenance
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15. In the Configure Magnification dialog, click the Setting drop-down and select the objective
with the next highest numerical aperture (NA) that is in position #2.
Repeat the following steps for each objective and write down the focus values for each.
You will use these values to determine the parfocality for each objective. This is different
than configuring Z offsets for laser autofocus.
Note: You must use the same filter set for all objectives.
16. In the Move Stage to Absolute Position dialog, use the Current Position: Z control to find
and focus a sample in the live image window.
Since you found the X and Y coordinates of the sample for the first objective, you need to
change only the Z position for the other objectives.
17. When the sample is in focus, write down the Current Position: Z value.
18. After you write down the focus values for each objective, in the Plate Acquisition toolbar,
click , then click to move the stage to the load position.
19. Remove the bead plate, and then click again to return the stage to its previous
position.
20. Continue to update the software settings:
If the objective you change is the one you use to determine the plate bottom
reference point, see
Determining the Plate Bottom Reference Point After Changing
the Reference Objective on page 222 .
Whenever possible, leave the 10x objective in place and replace the other objectives.
If the objective is NOT the one you use to determine the plate bottom reference point,
exit the MetaXpress software and see
Entering the Focus Objective Values in the
Meta Imaging Series Administrator Software on page 228 .