Acquiring image series
5.5. Acquiring a Z-stack
With the
Z-Stack
acquisition process, you acquire a series of frames one after
the other, a Z-stack.
Acquiring a Z-stack
Example: You want to acquire a Z-stack. The sample is approximately 50 µm
thick. The Z-distance between two frames is to be 2 µm.
1. Switch to the "Acquisition" layout. To do this, use, e.g., the
View > Layout >
Acquisition
command.
2. On
the
Microscope Control
toolbar, click the button with the objective that
you want to use for the image acquisition.
Selecting the objective
3. Switch to the live mode, and select the optimal settings for your acquisition,
in the
Camera Control
tool window. Pay special attention to setting the
correct exposure time. This exposure time will be used for all of the frames in
the Z-stack.
Setting the image
quality
4. Search out the required position in the sample.
5. Activate
the
Process Manager
tool window.
6. Select
the
Automatic Processes
option.
Selecting the
acquisition process
7. Click
the
Z-Stack
button.
The button will appear clicked. You can recognize this status by the
button's colored background.
The
[
Z
] group
will be automatically displayed in the tool window.
8. Should another acquisition process be active, e.g.,
Multi Channel
, click the
button to switch off the acquisition process.
The group with the various acquisition processes should now look like
this:
9. Select
the
Range
entry in the
Define
list.
10. Enter the Z-range you want, in the
Range
field. In this example, enter a little
more than the sample's thickness (= 50 µm), e.g., the value 60.
Selecting the
acquisition parameters
11. In the
Step Size
field, enter the required Z-distance, e.g., the value 2, for a Z-
distance of 2 µm. The value should correspond with your objective's depth of
focus.
In
the
Z-Slices
field you will then be shown how many frames are to be
acquired. In this example, 31 frames will be acquired.
12. Find the segment of the sample that interests you and focus on it. To do this,
use the arrow buttons in the [
Z
] group. The buttons with a double arrow
move the stage in larger steps.
13. Click the
Start
button.
Acquiring an image
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