Leica TCS SP8 SMD, User Manual

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135
FLIM Data Acquisition
Whenever you toggle between these options, the split-off filter on the stand and the X1 port
are automatically moved to the correct position and the corresponding detectors are
activated.
If there are pulsed diode lasers in your system, this selection also impacts the automatic
adjustment of the fluorifier disc (if present), according to
Table 19, page 143 .
The value displayed in the
Count Rate Monitor
is the count rate of the automatically selected
detectors (thus the APDs for external FLIM, the SP FLIM detectors for internal FLIM, and the
HyD RLD for NDD FLIM). Detectors that have been activated but not used display a dark
count rate, whereas detectors that have not been switched on or activated display the
number
0
.
You can change the automatically applied settings later.
FLIM data is collected on the separate SMD workstation using the SymPhoTime software
and the FLIM image is displayed there. In addition, if APDs or SP FLIM detectors are used,
an intensity image is generated in the LAS AF software during the collection of FLIM data.
16.2.1.1 FLIM Data Acquisition with Internal SP FLIM Detectors
For a 5-channel system with a single internal SP FLIM detector, the detector should be at
position 4; two internal FLIM detectors should occupy positions 2 and 4 (
Figure 120 ). These
settings have been preconfigured and cannot be changed by the user. SP FLIM detectors
can be used for normal intensity image acquisition (
Chapter 16.1.1 ) as well as FLIM data
acquisition.
Figure 120: Beam Path Settings in LAS AF for internal FLIM data acquisition. Other detectors may also be
activated at the same time.
Proceed as follows to prepare the TCS SP8 for the collection of SP FLIM data:
1. Open the Setup tab.
2. In the FLIM Detector pull-down menu, select the Internal (SP FLIM) option ( Figure 119 ).
The SP FLIM detector should be automatically activated in the FLIM mode.
If the SP FLIM detector has been deactivated by the user in the meantime, it can be
activated by clicking the
ON button ( Figure 120, item 1 ).
Then the FLIM detector goes into FLIM mode. A Gain of 990 V suitable for FLIM data