Zeiss LSM 880, Руководство по эксплуатации

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CHAPTER 1 - SYSTEM OPERATION
ZEISS Left Tool Area and Hardware Control Tools LSM 880
146 000000-2071-464 10/2014 V_01
5.2.15.3 Performing a Z-Stack Using a
Z-Piezo
A Z-Piezo allows Z-Stacks to be acquired
considerably quicker compared to the focus drive
of the microscope stand. The speed advantage
depends on the total travel range in Z and the
speed of the acquisition of the individual image of
a Z-Stack. The available travel range of a Z-Piezo
device is much smaller compared to the Z drive of
the microscope. A Piezo device allows performing
smaller steps in Z with a higher absolute precision
for the individual steps. A Piezo device is
recommended for 3D measurements of objects
and for 3D superresolution imaging (Airyscan).
Tick the
Use Piezo
checkbox (only available if a Z-Piezo device is attached) to activate the usage of the
Z-Piezo device in the First/Last or Center modes (Fig. 195). In the SIM imaging mode this is the
obligatory mode and hence the checkbox is permanently ticked.
Clicking on the
Leveling
button in the
Focus
tool moves the Z-Piezo stage to its center position in
respect to the current focus position of the microscope focus drive. This function is used to set defined
initial conditions. The total Range of the Z-Stack is limited depending on the type of Piezo. Hence form
the center position the Z-Piezo stage can move half the total range each up and down.
Define the first and last slice using
First/Last
mode by pressing the
Set First
and
Set Last
buttons at the
respective focus positions. Adjusting the value for Number of Slices or Interval is independent form each
other and affects the Range of the stack. The Focus Position is not changed.
When changes are made to
Interval or Number of Slices in First/Last mode, the values are taken over
for the
Piezo
mode provided they are within the Z-Piezo stage focus work range. This also works in the
opposite direction.
The direction for acquiring the Z-Stack using the
Z-Piezo can be set by pressing the double-arrow button
(bidirectional scan) or single-arrow button (unidirectional scan). This function is only available in the SIM,
TIRF an LSM imaging modes. For bidirectional scan data acquisition is done also when the focus is moving
backwards to the initial start position of the stack. This only applies, however, if multiple stacks are
acquired in a time series. In unidirectional mode no data acquisition takes place during backwards
movement to the initial position of the stack in this case. If only one stack is acquired the direction
settings have no effect and data recording will always be unidirectional. A
Corr Z
input box is available
(only in SIM , TIRF and LSM imaging modes) to correct for possible shifts of the stacks in Z using
bidirectional mode.
Fig. 195 Use Z-Piezo for Z-Stack acquisition –
appearance for the LSM imaging
mode