Leica TCS SP8 SMD, User Manual

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FLIM Data Acquisition
Figure 138: Defining a single FLIM image
7. In the
Defining FLIM Image acquisition time
dialog, you can define the duration of the
acquisition of the individual image ( Figure 138 ). To improve FLIM data statistics, multiple
scanning operations are normally carried out with the specimen and the data is compiled
into a final image. The following alternative options are possible:
•
Duration of each image
: You can enter how long the data acquisition of an individual
FLIM image is to last. The corresponding number of scanning operations are carried
out on the specimen. The entered duration is corrected automatically so that a
scanning operation is always stopped at the end of an image.
•
Repetitions
: Enter the number of scans to be carried out.
• Acquire until max ___ photons/pixel : With this option, the data acquisition is not
stopped at a fixed point in time. Image acquisition ends if the brightest pixel contains
more photons than specified here. With this option, images with comparable
brightness and therefore comparable photon statistics can be acquired.
8. Start FLIM measurement by clicking on the image acquisition button Run FLIM .
In LAS AF, an image averaged across the entire measurement duration is shown.
16.3.4 Defining an xyz or xzy FLIM Stack
To acquire an FLIM stack, proceed as follows:
1. Select the specimen position.
2. In the Setup FLIM step on the Acquisition tab, select either the Acquisition Mode xyz or
xzy
as the scan mode (
Figure 137
). Optimize the instrument parameter setting for FLIM
measurement.
3. Go to the
Measurements
step in the FLIM Wizard.
4. Define a file name (see Chapter 16.3.2 ) in the Setup tab.
5. Open the
Acquisition
tab.