CHAPTER 1 - SYSTEM OPERATIONZEISS Left Tool Area and Hardware Control Tools LSM 880142 000000-2071-464 10/2014 V_015.2.15.1 Performing a Z-Stack UsingFirst/Last ModePress the First/Last mode button to open theFirst/Last definition panel.Press the Continuous action button and focusonto the specimen.While scanning change the focus using theAcquisition Parameter - Focus tool, the Z-StackNavigation (Fig. 193) or use the focus drive of themicroscope.When you reach the position within the specimen,where you would like to start the stack, hit SetFirst and this position (in μm referring to the 0position of the Z drive) will be taken as the firstposition in the stack.Move the focus into the other direction until theend position within the stack is reached, then hitSet Last. These focus positions are then indicatednext to Set First and Set Last buttons. The Rangedisplay box shows the total height of the stack inmicrometers.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 automatically adjusts the Interval in a way that thetotal Range for the stack is maintained.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 keeping the total Range for the stack within the possibilities of fitting the calculatednumber of slices into the Range. The total Range may therefore vary to higher and lower numbersaround the initial value of the Range.Pressing the Optimal button (only available in SIM and LSM imaging modes, greyed out in WF andLaser WF imaging modes) sets Interval to match the Nyquist criteria (2 fold over sampling) andaccordingly adjusts the Slices to keep the Range within the limits defined by Set First and Set Last. Theoptimal 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.Fig. 193 Z-Stack Navigation appearance forthe LSM image mode
