Fit Fluorescence Decays

The actions performed for each channel are:

• Load IRF data corresponding to the channel (*)

• Store resulting nanotime histogram as Donor IRF (**)

• Load channel data and perform a complete Data Analysis script

• Compute the nanotime histogram corresponding to the selected bursts resulting from the previous analysis

• Store this histogram as the D-only F^D fluorescence decay curve (**)

• Perform an unweighted fit of the decays using the current fit parameters as initial guess

• Perform a weighted fit using the previous fit parameters as initial guess

For each step, the options selected by the user are used.

For instance, the fitting parameter uncertainties will be computed and exported to the Notebook if and only if the Automatically Compute Parameter Uncertainties and Automatically Export Results to Notebook options have been selected.

Additionally, if the Generate HTML Report checkbox on the Multispot Analysis page is checked, the Nanotime Analysis graphs and corresponding fit results are exported as an HTML page, which is automatically opened in the default browser.

The list of actions described above are perforrned successively on each channel belonging to the following list:

• if All Channels is selected, all channels stored in the file are used

• if All Channels is not selected, only the channels selected on the Data File page are used.

Note that if the latter option is used, selecting more than one channel as a Donor will result in a warning of channel duplication (message and red LED on at the bottom of the ALiX window. This can be safely ignored, as the script will eventually process each channel separately.

(*) The IRF file used for the script needs to be specified in the IRF File Path box.

(**) If the Sum Channels checkbox is checked, the IRF and nanotime histogram will be added to the existing IRF and nanotime histogram (instead of replacing them).