Making asymmetric stacks in Mark & Find mode using Define Asymmetry

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1. Mark your first position (position 1). It doesn’t seem to matter whether you set up the Z series now or mark the rest of your points first. If ‘Same stack for all’ is checked, then the same asymmetries will be applied to all positions, so make sure the marked points are at similar focus positions.
2. Once you’ve set all your points return to position 1.
3. Expand the Z-Stack controls. Click the Set Plane button and select either Set Plane or Set Focus Plane. It doesn’t seem to matter which one you choose so I am only going to refer to the Focus Plane from now on.
4. Set the Begin and End of your Z stack.
5. Click the ‘Go to’ button. You can now toggle between the Center of your Z-Stack, which will be equidistant between the Begin and End, and the Focus Plane, which will be the same as the focus of position 1. You MUST go to the Focus Plane before returning to Mark & Find and setting the asymmetry, otherwise the Focus Plane will not be the plane used to define the asymmetry.
6. Click on Mark & Find again and IMMEDIATELY click Define Asymmetry. Do not explore the various points you’ve marked first, otherwise the link between position 1 and the center of the Z-Stack is lost.
7. You can now Start the acquisition.