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ELYRA 7
Left Tool Area and Hardware Control Tools
ZEISS
03/2019 V_02
000000-2262-999
129
5.2.17.3
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.
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. 198). In the
SIM Apotome
&
Lattice
SIM
imaging modes 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.
In order to acquire an extended 3D SMLM data set, set the number of slices and the z-range using
First/Last
or
Center
mode. Choose
Laser WF
imaging mode in the
Imaging Setup
tab and choose
from the
Trigger in
drop down menu of the
Acquisition
tab the entry
internal
. In this way the camera
exposure is triggered to the slice.
Please note that the software allows only a Z-Stack (inner loop) within a time series (outer loop),
but no time series of a Z-Stack. This means that for each time point of the 3D SMLM experiment,
only one image per slice is taken. It is not possible to take many images per slice before advancing
to the next slice. As this procedure will be time consuming, it is advised to keep the number of
slices to a minimum.
Fig. 198
Use Z-Piezo for Z-Stack acquisition