...
Open Mark & Find and click on the Stage Configuration button indicated by the red arrow (A) in the image below.
In the Stage Configuration window , that appears click the Symmetric radio button indicated by the red arrow in the image below(B).
| Note |
|---|
Always check that Mark & Find is in Symmetric mode before starting. Likewise, if you need to make asymmetric stacks you need to make sure it is set correctly for that too |
Close the Stage Configuration window and focus on the specimen plane that you want to be in the middle of the stack. This might be different to from the plane with the most fluorescence. For instance, an adherent cell may have be more fluorescence fluorescenct near the cover-slip where the cell is flattest, but the middle of the stack might be half way up the nucleus. It might be easiest to use a nuclear stain to locate the middle in this case but you will have to make sure the Begin and End of the stack are beyond the edges of the apical and basal membrane. Mark the position by clicking the button indicated by the arrow belowC above. Mark Make sure 'Same stack for all' is checked.
| Info |
|---|
Note that the 'Define Asymmetry' button in the bottom right of the image below will not appear when Mark & Find is in Symmetric mode |
Set up a Z series around the point you marked. The Begin and End of your series will automatically adjust so the marked point is the centre of the stack. Make sure the Z Step is set correctly. Mark more points; they will all automatically be the centre of Z stacks of the same range and Z Step as the first point.
...