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Repetition rate with pulsed diode lasers
In the supplied configuration, 40 MHz is the maximum repetition rate for
the pulsed diode lasers. 80 MHz is available only if the trigger on the laser
driver (PDL, SEPIA) is set to internal. This bypasses the laser settings in
LAS AF. In addition, all functions related to beam blanking (such as ROI
scanning or line-by-line sequential scanning) are deactivated. A change
of synchronization cable length will be necessary as well.
1. Run a test measurement by selecting Run FLIM Test in LAS AF.
2. The Measurement Preview window opens in SymPhoTime (Figure 148 right). Close this
window by selecting the Stop button.
3. You can now test the fading behavior of your specimen using the oscilloscope window
in the SymPhoTime software. Select Show Oscilloscope...
4. Select the TCSPC tab Histogram, select Start and monitor the fading of the fluorescence
as shown in Figure 149.
Figure 149:
Fading of the fluorescence at too high of a laser repetition rate (A, 80 MHz), optimum laser
repetition rate (B, 40 MHz) and unnecessarily low laser repetition rate (C, 20 MHz).
5. In this window, you can select the time resolution per bin of the TCSPC histogram with
the slider under Acquisition.
6. Please confirm your selection by selecting Apply.
Summarized manual for FLIM experiments
p
169
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