1. The EDF (motorized focus) workbench is selected.
  2. The sample is sufficiently illuminated and in focus.
  1. Set up the camera using the Camera tool.
  2. Set up the focus range over which to acquire the EDF image in the Motorized Extended Depth of Focus tool.
  3. Move the stage away from the sample until the top of the sample is no longer in focus. Define this focal plane position as the upper end of the range by clicking Top > Assign.
  4. Move the stage towards the sample until the bottom of the sample is no longer in focus. Define this focal plane position as the lower end of the range by clicking Bottom > Assign.
  5. The step size is calculated automatically. If you want another step size, set the value for the Step Size to specify the distance the stage travels between two acquisitions.
  6. The number of images to be acquired is displayed automatically as Slices.
  7. Alternatively you can use Optimal: The optimal Step Size is determined automatically depending on your microscope setup. The step size is calculated according to the Nyquist criteria.
  8. Start the acquisition by clicking the Start button in the Workbench Area.

acquires the slices and calculates the EDF image automatically.

1

This position or slightly higher: Top > Assign

2

This position or slightly lower: Bottom > Assign

The algorithm used to calculate the EDF image works best if the acquired images differ significantly. Try to acquire as few images as possible. Make sure an acquired image contains focused areas which are not focused in the other images, i.e. each new image contains new information relevant for the EDF calculation.