Lightfield 4D experiments support the use of different focus strategies:

• Software Autofocus: The focus is found by scanning a predefined volume with a reference channel that has to be part of the track setup (the reference channel can be deactivated). Note that the Lightfield 4D channel itself cannot be used as a reference for Software Autofocus! For Lightfield 4D a valid scenario would be an activated Lightfield 4D track combined with a deactivated confocal track as reference channel. Use the confocal track for finding focus with autofocus. The software autofocus is not supported for setups without any reference channel.
• Absolute Fixed Z Position for the whole experiment. The position is either set manually or derived from z-positions defined from a multi-position experiment.
• Definite Focus: The Definite Focus device can run independently of the acquisition but needs specific conditions to work, like a glass-water or air-glass interface to find the reflex of the in build red diode which is projected onto the sample and the reflex is then registered by the attached detector. Note that for high-speed acquisition the Definite Focus is only executed before the time series starts and never during the fast acquisition.