Leica TCS SP8 SMD, User Manual, Page: 154 / 270

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FLIM Data Acquisition
6. Define the first and last image of the stack and the number of planes via the stack dialog
( z-stack or y-stack ) ( Figure 139 ).
Figure 139: FLIM stack definition xzy
7. Define the duration of the individual FLIM images as described in Chapter 16.3.3 . The
option Acquire until max ___ photons/pixel is of particular interest for acquiring FLIM
stacks. It enables corrections due to intensity changes caused by different sample
penetration depths.
8. Start the FLIM stack acquisition with the image acquisition button Run FLIM .
In LAS AF, you obtain a stack of averaged images and, in SymPhoTime, a series of FLIM
images.
16.3.5 Defining an FLIM Time Series
To acquire an FLIM stack, proceed as follows:
1. Select the specimen position.
2. Select either Acquisition Mode xyt or xzt as the scan mode on the Acquisition tab.
Optimize the instrument parameter setting for FLIM measurement.
3. Define a file name in the Setup tab ( Chapter 16.3.2 ).
4. Open the Acquisition tab.