Leica TCS SP8 SMD, User Manual

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FLIM Data Acquisition
5. Define the duration of the individual FLIM images as described in
Chapter 16.3.3
. For
recording a FLIM time series, use either Duration of each image or Repetitions , because
the last option, (
Acquire until max ___ photons/pixel
), does not allow a fixed period of
time to be specified.
6. You can define a time series using the time series dialog (
Figure 140
). The following
inputs are possible:
•
Time Interval
: Defines the time between the beginning of one FLIM image and the
next image. This duration cannot be shorter than the duration entered for a single
FLIM image. To specify the smallest possible duration, select
Minimize
.
• Duration : Here, enter how long the entire time series is to last. The corresponding
number of FLIM images is acquired. The duration entered is automatically corrected
so that all FLIM images can be completed.
•
Frames
: You can specify the number of FLIM images to be acquired. The maximum
specified number is 1000.
Figure 140: Defining an FLIM time series
7. Enter the desired value in the time series dialog.
8. Start the FLIM time series acquisition with the image acquisition button
Run FLIM
.
In LAS AF, you obtain a time series of averaged images and, in SymPhoTime, a series of
FLIM images.
16.3.6 Defining a Time Series of xyz or xzy FLIM Stacks
Proceed as follows to take a time series of FLIM stacks:
1. Select the specimen position.
2. Select either
Acquisition Mode xzyt
or
xyzt
as the scan mode on the
Acquisition
tab.
Optimize the instrument parameter setting for FLIM measurement.