ELYRA 7
Left Tool Area and Hardware Control Tools
ZEISS
03/2019 V_02
000000-2262-999
125
5.2.17.1
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. 196) 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.
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
µ
m
. 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
(Opt)/(Leap)
button (only available in
SIM Apotome and Lattice SIM
imaging mode, you
can select between Optimal (Nyquist) and Leap (3 fold broader spacing)).
Optimal
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
. Leap
sets the spacing 3 fold broader than Nyquist.
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
.
The
Position (
µ
m)
input box indicates the current position of the focus, no matter if the focus drive or a
Z-Piezo
stage insert was used for positioning. The focus is changed in small steps when clicking the
arrow keys.
If the
Use Piezo
check box is ticked (only available, if a
Z-Piezo
stage insert 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 drive was used for
Set First
/
Set Last
, the system will transform this numbers for a matched
Z-
Piezo
movement. In the
SIM
imaging mode, the
Use Piezo
check box is ticked by default and greyed
out, so there is no choice to deselect the box and use the microscope focus drive for
SIM
imaging.
Fig. 196
Z-Stack Navigation appearance for
the SIM Apotome and Lattice SIM
image modes