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In the ND Acquisition window tick the Save to File box and define a filename and path for the timelapse . Tick ‘Close active shutter during Z movement’ to reduce photobleaching(see below).
Click the lambda tab and select the channels you want to image by ticking the box at the start of each row and selecting from the drop-down list of optical configurations (red arrow below). Tick the box at the bottom of the window that says ‘Close active Shutter during Filter Change’ if you want to reduce photobleaching.
Click the XY tab and click the Add button to add points as you move the stage around. Make sure ‘Close active Shutter during Stage Movement’ is ticked to reduce photobleaching and that the ‘Include Z’ box is ticked so the Z position is included along with the XY position. If the PFS is switched on the offset will be saved in the PFS column along with the X, Y and Z value at each position. It may be necessary to adjust the PFS offset at different positions, especially when marking points that are very distant to one another. If you mark a position with the PFS off and the focus changes when you turn the PFS on then the offset is different at that position. You can adjust it using the wheel on the PFS Offset Controller and then update the value in the software by clicking the arrow next to the PFS column. X, Y and Z positions can be updated in the same way. Finally, marked positions can be reordered using the up and down arrows in the toolbar and pressing the Optimize button will re-order the points so that the stage moves the shortest distance.
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