Leica TCS SP8 SMD, User Manual

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FLIM Data Acquisition
10. Define the duration of the individual FLIM images as described in
Chapter 16.3.3
. If all
images are to have a comparable brightness, use the Acquire until max ___ photons/
pixel
option. It enables corrections of intensity changes which are caused by the
excitation spectrum and by bleaching artefacts. If intensity information is required, use
Duration of each image
or
Repetitions
.
11. Start the FLIM stack acquisition with the image acquisition button Run FLIM .
In LAS AF, you obtain a
Λ
stack of time-averaged images and, in SymPhoTime, a series of
FLIM images.
16.3.10 Control of FLIM Measurements
The measurement series is started by clicking on the
Start FLIM
image acquisition button. It
stops automatically after all measurements are taken. A user-defined stop is possible by
clicking the
Stop FLIM
image acquisition button.
In the count rate monitor ( Figure 147 ), the current count rate (in cps - counts per second) is
displayed during the individual measurements. In the line Max. counts per pixel , the total
number of photons accumulated so far in the brightest pixel of the image is shown. This
gives an initial indication as to whether the photon statistics in the image are sufficient.
After start of the FLIM series, fast FLIM images are displayed in SymPhoTime online and
saved together with the raw data.
Both the count rate and fast FLIM images give a first impression about the quality of the
measurements.
Figure 147: Count rate monitor in the Measurements step of the FLIM Wizard