CHAPTER 1 - SYSTEM OPERATIONZEISS Left Tool Area and Hardware Control Tools LSM 880144 000000-2071-464 10/2014 V_015.2.15.2 Performing a Z-Stack Using CenterModePress the Center mode button to open the Centerdefinition panel (Fig. 194). This mode allowssetting the Range of the Z-Stack in relation to thecurrent Focus Position which is indicated asOffset.In addition it is possible in the LSM imagingmode to use the image of a XZ scan (line scan inZ) for positioning the Z-Stack range. This tool setsthe first and last slice symmetrically in relation to adefined Center position. One should have an ideaabout the size of the stack when using this mode.It is useful when several Z-Stacks of the samerange should be acquired at different positionswithin the specimen, as the whole range of thestack can be set to a different position at onceusing the Offset function.The values for number of Slices and Interval are taken over from any settings done in First/Last mode.The Offset defines the Center position of the stack in relation to the Focus Position.The Slices input box displays the number of slices of the stack. This number can be edited by typing orusing the arrow keys. Changing the value for Slices leaves the Interval untouched, but the total Rangeis adjusted.The Interval input box displays the z-distance (interval) between slices in μm. This number can be editedby typing or using the arrow keys. Changing the value for the Interval automatically adjusts the numberof Slices while adjusting the total Range. The Slices number will remain unchanged.Pressing the Optimal button sets Interval to match the Nyquist criteria (2 fold over sampling) andaccordingly sets the number of Slices. The optimal interval is displayed in the button in μm.Within the Keep display field (only available in Show all mode) you have the option to switch betweenKeep Interval and Keep Slice. If Keep Interval is checked, the number of slices is adjusted to a newRange defined pressing Set First or Set Last. Likewise, when Keep Slice is checked, the value for theInterval between the slices is adjusted to a new Range defined by pressing Set First or Set Last.The Position (μm) input box indicates the current position of the focus, no matter if the focus drive or aZ-Piezo stage insert was used for positioning. The arrow keys allow to change the focus in small steps. Ifthe Use Piezo check box is ticked (only available, if a Z-Piezo stage is attached to the system), the Z-Piezo stage insert will be used to perform all Z movements triggered from the software. If the focus drivewas used for Set First / Set Last, the system will transform this numbers for a matched Z-Piezomovement.The Fast Z Line checkbox is only available in LSM imaging mode and only Line Scan Mode to beselected in the Acquisition Mode tool. It is not possible with the Z-Piezo. If the check box is ticked andContinuous action button pressed a fast XZ scan for overviews is performed using a continuousmovement of the microscope focus. The stack size is retained and the interval is adapted depending onthe scan speed.Fig. 194 Z-Stack tool Center mode withZ-Stack Navigation displayed forthe LSM image mode
