*For a single immobilized molecule antibunching trace you will need at least a trace of 30s fluorescence emission with a minimum mean countrate of 10 kcps.
alhistograms.png | Original data file content }}
Trace #0 shows the IRF, Trace #2 is the decay curve. The other data blocks are empty. Both Trace #0 and #2 will be imported into a single ''.etc''... n order to perform reconvolution fitting of the Antracene decay, the corresponding IRF measured at 375nm
ult-file opens with a preview display (Image, FCS-traces, TCSPC-curve or Time Trace - depending on the type of analysis). Additionally the **"Comment"** - tab o... of the crosscorrelation is calculated.
=== Time-Trace ===
* Max. Counts
* Av. Counts
=== TCSPC ===
on SymPhoTime}}
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====== Calculate FCCS Traces with the Grouped FCS Script ======
===== Summar... ere.\\
- **Upper center/right:** Intensity time trace. The display option can be changed using the "Trace Settings" of the analysis options. The large window shows the inset of the complete trace above marked in green. The photon counting histog
age.\\
- **Upper center/right:** Intensity time trace. The display can be changed using the "Trace Settings" of the analysis options. The large window shows the inset of the complete trace above highlighted in green. The photon counting h... of the different intensity values. Usually, this trace is used to check, whether the signal is stable du
nter, the count rate should stay stable. The time trace in figure 8 shows a reduction of the countrate du... , Inhouse II, MPD/SPAD}}]
[{{ howto:diamond_timetrace.png?270x200|Figure8: No bleaching visible. reduct
{{tag> tutorial howto time-trace analysis FRET SymPhoTime}}
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====== Calculate Ratiometric Singl... requires a spectrally resolved fluorescence time trace. As a result, the distribution of FRET efficienci... sites =====
**Note:** The script requires a time trace containing the fluorescence of two spectrally sep... the fluorescence of the acceptor dye.\\
The time traces can be acquired from single molecule events in a
{{tag> tutorial howto FRET PIE analysis time-trace SymPhoTime}}
~~NOTOC~~
====== Calculate Ratiometric S... requires a spectrally resolved fluorescence time trace, in which donor and acceptor have been excited al... rY?large }}
**Note:** The script requires a time trace containing the fluorescence of two spectrally sep... the fluorescence of the acceptor dye.\\
The time traces can be acquired from single molecule events in a
oTime}}
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====== Calculate and Fit FCS Traces with the FCS Script ======
===== Summary =====
... ide of the main window.
{{ calculate_and_fit_fcs_traces_with_the_fcs_script_Image_1.png }}
* Highligh... y a single mouse click.
{{ calculate_and_fit_fcs_traces_with_the_fcs_script_Image_2.png }}
* Select t... he dropdown menu "FCS".
{{ calculate_and_fit_fcs_traces_with_the_fcs_script_Image_3.png }}
**Note:** Th
ile up limit the count rate displayed in the Time Trace below the FLIM image is used.
* A red line indi... 0% of the laser repetition rate.
* The **"Time Trace Binning"** should be set to 0.01 ms which equals ... he Leica SP8 SMD. Here, the bin width in the time trace is fixed at 1 msec averaging many pixels. This mi... each pixel is displayed as one point in the time trace. At a pixel dwell time of 1 µs the average of 10
steps according to the tutorial [[Calculate FCCS Trace with the Grouped FCS Script]].
**Note:** This wa... page.
- **Upper center/right:** Intensity time trace. The display can be changed using the "trace display settings of the analysis options. The large window shows the inset of the complete trace above highlighted in green. The photon counting h
{{tag> tutorial howto time-trace analysis SymPhoTime}}
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======Intensity Time Trace Analysis======
===== Summary =====
This tutor... al shows step-by-step, how to draw intensity time traces from a single molecule. The script requires a po... of the main window.
{{ using_the_intensity_time_trace_script_Image_1.png }}
* Highlight the file ''C