LEFT TOOL AREA AND HARDWARE CONTROL TOOLS
LSM 710 and LSM 780
Carl Zeiss
Acquisition Tab
Systems
88 M60-1-0025
e
02/2010
that position. A Z-Stack is then acquired at the new position when starting the experiment. Hitting
Center
moves the
Center
position back to the actual
Focus Position
setting the
Offset
again to zero.
Use
,
and
to get images from these three specific positions. The
Focus Position
updates accordingly. The
Offset
value then might be set to half of the Interval depending on the Interval
value and the number of slices, as with a lower number of slices or a large Interval the real center
position of the stack might be just between two slices.
Range Select
produces a XZ-scan within the
Range
of the Z-Stack. This helps setting the
Range
of the
Z-Stack without continuously scanning complete frames. The position of the first and last slice is indicated
with a red line, the
Focus position
, which initially corresponds to the Center position, is indicated as a
green line.
Changing the
Offset
value moves the red lines in parallel to the new offset position, the green line, the
Focus Position
is kept constant.
sets the
Focus Position
to the indicated
Center
position setting
the
Offset
back to zero and producing an image at the newly defined Focus Position. Alternatively
hitting Center sets the indicated Center position back to the actual Focus Position moving the red lines
back to the original position and setting the Offset back to Zero. The original values for Number of Slices,
Interval and Range are always kept constant.
The red lines can be moved independently from each other allowing an optimal adjustment of the
Z-Stack range to the specimen.
Center
sets the green line to the center position between the red lines
and adjusts the Focus Position accordingly. The original values for
Number of Slices
and
Range
are
adjusted. The
Interval
is kept constant.
The size and position of the Line where the XY scan should be performed can first be defined
using the line scan function in the Acquisition Mode tool.
5.2.12.3
How to Proceed Setting a Z-Stack
Using Piezo Mode
The Piezo objective focusing device allows stacks to
be produced considerably quicker than via the
focus of the microscope stand. Clicking on the
Leveling
button in the
Focus
tool moves the Piezo
objective focus to the zero position, while the
microscope focus moves into the opposite
direction at the same time, i.e. the position of the
object in relation to the objective remains
unchanged. This function is used to set defined
initial conditions.
•
Define the first and last slice using
Start/End
mode. Activate
Piezo
mode (Fig. 78). The total
Range of the Z-Stack is limited to 250 µm when
using the Piezo device. 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
Start/End mode
, the values are taken over
for the
Piezo
mode provided they are within the Piezo objective focus work range. This also works in the
opposite direction.
Fig. 78
Z-Stack tool Piezo mode with Z-Stack
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