Leica TCS SP8 SMD, User Manual

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FLIM Data Acquisition
16.2 Setup FLIM Step – Optimizing the FLIM Measurement
Conditions
Go to the Setup FLIM step in the FLIM Wizard. Now set the instrument parameters for a FLIM
measurement.
16.2.1 Selecting FLIM Detectors
For FLIM measurements, special detectors have to be used that are able to do single photon
counting with a high time resolution. With FLIM excitation in the UV and VIS range, FLIM
detectors are used in a descanned position: The light is either split off at the X1 port to APDs
or a FLIM photomultiplier (external detection) or sent to the SP detection module on the SP
FLIM photomultiplier (internal detection). With MP excitation, FLIM detectors may optionally
be used at a non-descanned position. When doing so, the fluorescent light on the stand is
split off directly behind the objective. The following options are available:
• External FLIM detection using two APDs
• External FLIM detection using a FLIM PMT
• Internal FLIM detection using one or two SP FLIM PMTs
• External FLIM detection using two APDs and internal FLIM detection using two SP FLIM
PMTs; the internal and external detectors can only be used in alternation for this
• Detection at NDD position with one or two HyD RLD
If you use a system with both internal and external FLIM detection, first you have to select
the detectors you want to use:
1. Select the
Setup
tab (
Figure 119
).
2. In the FLIM Detector pull-down menu, select either Internal (SP FLIM) for internal SP
FLIM detection,
External
for external detection at the X1 port, or
Non-descanned (MP
FLIM) for detection at an NDD position ( Figure 119 ).
Figure 119: Setup FLIM step