Leica TCS SP8 SMD, Руководство пользователя, Страница: 160 / 270

160
FLIM Data Acquisition
9. Start the FLIM
λ
stack acquisition with the image acquisition button
Run FLIM
.
In LAS AF, you obtain a
λ
stack of averaged images and, in SymPhoTime, a series of FLIM
images.
16.3.9 Defining an xy
Λ
or xz
Λ
FLIM Stack
These modes are only available for systems that are equipped with variable-frequency
lasers (white light lasers, MP lasers). In the following, the acquisition of xy or xz FLIM
stacks with the white light laser is described:
1. Select the specimen position.
2. In Beam Path Settings in the white light laser dialog, select the mode Constant Power
(
Figure 144
).
Figure 144: Selecting the Constant Power mode
3. In the Setup FLIM step on the Setup tab, select the option Internal (SP FLIM) or External
(
Chapter 16.2.1
).
4. In the Measurements step on the Acquisition tab, select either the Acquisition Mode
xy
Λ
or
xz
Λ
as the scan mode.
5. Check that the correct detector for the FLIM measurement is activated in the
Λ
stack
dialog.
6. Optimize the instrument parameter setting for FLIM measurement. While doing so, test
different excitation wavelengths.
7. Define a file name in the Setup tab ( Chapter 16.3.2 ).
8. Open the
Acquisition
tab. If you want to use a spectral internal FLIM detector, select it
in the PMT Selection field (Figure 146 ). If you remove the check next to Automatic SP
Movement
, you can manually select the external detector manually with white light or
MP laser excitation in Beam Path Settings .
Λ Λ