Performing A Z-Stack Using First/Last Mode - Zeiss LSM 880 Operating Manual

ZEISS
Fig. 193 Z-Stack Navigation appearance for
the LSM image mode
The Slices input box displays the number of slices of the stack. This number can be edited by typing or
using the arrow keys. Changing the value for Slices automatically adjusts the Interval in a way that the
total Range for the stack is maintained.
The Interval input box displays the z-distance (interval) between slices in µm. This number can be edited
by typing or using the arrow keys. Changing the value for the Interval automatically adjusts the Number
of Slices while keeping the total Range for the stack within the possibilities of fitting the calculated
number of slices into the Range. The total Range may therefore vary to higher and lower numbers
around the initial value of the Range.
Pressing the Optimal button (only available in SIM and LSM imaging modes, greyed out in WF and
Laser WF imaging modes) sets Interval to match the Nyquist criteria (2 fold over sampling) and
accordingly adjusts the Slices to keep the Range within the limits defined by Set First and Set Last. 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 between
Keep Interval and Keep Slice. If Keep Interval is checked, the number of slices is adjusted to a new
Range defined pressing Set First or Set Last. Likewise, when Keep Slice is checked, the value for the
Interval between the slices is adjusted to a new Range defined by pressing Set First or Set Last.
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CHAPTER 1 - SYSTEM OPERATION
Left Tool Area and Hardware Control Tools
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5.2.15.1 Performing a Z-Stack Using
First/Last Mode
Press the First/Last mode button to open the
First/Last definition panel.
Press the Continuous action button and focus
onto the specimen.
While scanning change the focus using the
Acquisition Parameter - Focus tool, the Z-Stack
Navigation (Fig. 193) or use the focus drive of the
microscope.
When you reach the position within the specimen,
where you would like to start the stack, hit Set
First and this position (in µm referring to the 0
position of the Z drive) will be taken as the first
position in the stack.
Move the focus into the other direction until the
end position within the stack is reached, then hit
Set Last. These focus positions are then indicated
next to Set First and Set Last buttons. The Range
display box shows the total height of the stack in
micrometers.
LSM 880
10/2014 V_01