Leica TCS SP8 SMD, User Manual

Page: 164 / 270

164
Summarized manual for FLIM experiments
applies for pulsed diode lasers and white light lasers with a pulse picker only. The
repetition rate of multiphoton lasers cannot be adapted (other than with a pulse picker).
For details, see
Chapter 17.9
.
17.4 Changing from Internal Detection on the SP8 to External
TCSPC Detectors
1. In the FLIM Wizard in LAS AF, go to step
Setup FLIM
.
2. If you want to carry out your time-resolved experiments with the external PicoQuant
detectors, configure the detection beam path in the scan head to direct the fluorescence
to the X1 output port (that means the
X1 Port at position --- )(Chapter Chapter 16.2.1 and
Chapter 16.2.1.2
). This is done automatically when you select the
External
FLIM detector
in the " Setup " tab ( Chapter 16.2.1 , Figure 119 ).
3. If you have two single detectors, set the appropriate SMD filter cube inside the X1 port
adapter to split the appropriate spectral range of the emission light to the two detectors.
4. Make sure that suitable fluorescence detection filters (band pass / long pass) are
present in the corresponding filter holders of the PicoQuant detector unit(s). If the
detection filters and beam splitters are already placed inside the X1 port adapter, you
can leave the filter holders in front of the detectors empty. However, a filter holder must
be located in front of the detector in order to open its shutter.
17.5 Using Internal SP FLIM Detection
1. In the FLIM Wizard in LAS AF, go to step
Setup FLIM
.
2. Select the FLIM detector Internal (SP FLIM) on the Setup tab ( Chapter 16.2.1 , Figure 119 ).
This automatically moves the X1 port to the position
Mirror
that reflects the emission
light to the SP detection module within the SP8 scan head.
3. Select a suitable spectral detection range by moving the mirror sliders of the detectors
in the Beam Path Settings (Chapter Chapter 16.2.1.1 ).
4. Check whether the fluorifier disc is in the correct position (
Chapter 16.2.3
).
17.6 Using FLIM Detectors at the NDD Position (HyD RLD)
1. In the FLIM Wizard in LAS AF, go to step Setup FLIM .
2. Select the FLIM detector Non descanned (MP FLIM) on the Setup tab ( Chapter 16.2.1 ,
Figure 119 ). By doing so, a beam splitter in the stand which decouples the florescent light
to the HyD RLD automatically swings into the beam path.
3. Make sure that the correct barrier filter and the suitable filter cube are placed in the
Pay attention to the APD detection unit manual
When using a two-channel APD detector unit from PicoQuant, ensure
that the beam splitter in the beam splitter turret is set so that it distributes
the emission light to the detectors correctly. Detailed information can be
found in the detection unit manual.