Leica TCS SP8 SMD, User Manual

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Summarized manual for FCS or other point measurements
6. Select the measurement times.
7. Start the FCS measurement in LAS AF by selecting the image acquisition button Run FCS
under the step
Measurements
.
Normally, the SymPhoTime software will start the data acquisition automatically. If you
start the FCS data acquisition by selecting
Acquire Point Measurements
in the
SymPhoTime software, the measurement is not synchronized with LAS AF. The file
name, comments and instrument parameter settings are not transferred from LAS AF to
SymPhoTime.
8. The Measurement Preview dialog opens in SymPhoTime on the
FCS
tab (
Figure 182
,
left
).
The FCS curve is integrated over time.
Figure 183: Working area of the annotation file
9. At the end of the measurement, the result can be viewed in the annotation file which can
be opened from the workspace (
Figure 183
). Open the
Correlation
tab as shown in
Figure 184 . The curves can be analyzed by selecting the FIT button that corresponds to
the cross-correlation or autocorrelation curve.
10. Starting from the acquired raw data file (in this case, FCS 01.pt3 ), the FCS curve can be
recalculated with enhanced functions, such as Time Gated FCS or Fluorescence Lifetime
Correlation Spectroscopy (FLCS) using the
FCS trace button. The prerequisite for both of
these modes is that the FCS measurement was carried out using pulsed lasers for
excitation.