Hi µManager community!
I would like to submit a problem we are encountering regularly, for which we've been stuck for days now. We perform multi-position, 4D acquisitions (Z-stacks at several timepoints). We usually program ~10 positions and acquire ~20 Z planes in two channels. The total acquisition time is typically 2-8 hours.
To avoid axial drift, we activate the Nikon Perfect Focus system. It works well most of the time, and finds the focus at the beginning of almost every single Z-stacks. However, sometimes, it will not find the focus and will output the following error message in the logs:
In our case, we can afford having the PFS not finding the focus once in a while, but a major issue is that after this bug, the stage always stop scanning in z, and all the subsequent images are acquired with the stage stuck at the same Z position.
Is this an expected or at least documented behaviour? Would you have any advice about how to circumvent it? How could we recover Z-scanning after this issue?
Antoine Coulon lab, UMR1664
Institut Curie, 26 rue d'Ulm, Paris
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