[Vespera Pro] Does automatic calibration apply when capturing mosaics and using Plan my Night?

Yes. Ensure that the feature is enabled before starting the capture.

How does automatic calibration affect multi-night observations?

If you start a multi-night observation with automatic calibration enabled, it will remain active when resumed, regardless of the setting in the Singularity interface.
Conversely, if automatic calibration is disabled at the start, it will remain disabled for the resumed observation.

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