Leica TCS SP8 SMD, User Manual

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157
FLIM Data Acquisition
7. Enter the desired value in the time series dialog.
8. Start the FLIM experiment recording with the Run FLIM button.
In LAS AF, you obtain a time series of averaged images of stacks and, in SymPhoTime, a
series of FLIM images.
16.3.7 Defining an xy
λ
or xz
λ
FLIM Stack
These modes are only available for systems that are equipped with internal SP FLIM
detectors. During the generation of stacks, the spectral sliders of the SP FLIM detectors
are shifted incrementally. To acquire an FLIM stack, proceed as follows:
1. Select the specimen position.
2. In the Setup FLIM step on the Setup tab, select the Internal (SP FLIM) option
(
Chapter 16.2.1
).
3. In the Measurements step on the Acquisition tab, select either the Acquisition Mode
xy
λ
or
xz
λ
as the scan mode. Optimize the instrument parameter setting for FLIM
measurement.
4. Define a file name in the
Setup
tab (
Chapter 16.3.2
).
5. Open the Acquisition tab. If the correct SP FLIM detector is not yet selected, select it in
the
PMT-Selection
line. In
Beam Path Settings
, the detector selection cannot be
changed.
6. Define the stack in the
λ
stack dialog as follows (
Figure 142
):
• Detection Begin and Detection End : Wavelength of the first and last FLIM image
•
Detection Band Width
: Width of the spectral range of an individual FLIM image
• No. of Detection Steps : Number of FLIM images that are acquired from different
spectral ranges
• -Detection Stepsize : Step size of the spectral slider from one image to the next
λ
λ