This module offers a configurable image based autofocus functionality that will search through a series of axially stepped images analyzing the “sharpness” of each. The z-value of the image returning the maximum sharpness is set as the new plane of observation.

The module requires the microscope to be fitted with a motorized Z-drive. It does not require a z-piezo actuator nor is the z-piezo used by the software autofocus (SWAF) in the current implementation. On the Acquisition or Locate tab the settings for the SWAF can be adjusted in the Software Autofocus tool. These can also be called (and tested) by clicking the Find Focus (AF) button on the main button bar on Acquisition tab. SWAF settings are stored as part of an experiment on Acquisition tab.

The configuration allows the function of the SWAF run to be matched to the conditions under which the focus should be found. A basic description of the functions adjusted by the individual controls can be found further below. However, before going into the description of each parameter, we will try to address the following questions: How does the SWAF in the software attempt to locate the “focus”? And how do the parameters settings influence its behavior in this respect?