Embedded Files
The Lifetime Graph shown below is used for different purpose depending on the context, as described in the different sections below. Notice that the graph has two vertical axes. The left axis (Lifetime (ns))corresponds to lifetime (or in general time) values, while the right axis (Other Parameters) is used to plot fit parameters of decay series (see Fluorescence Decay Fitting manual page).
The horizontal axis (labeled Sample in the picture below) might take on a different name depending on context (Time (s) for instance, for a time series analysis). Axes labels can be edited in the Scale Legend box (between the Graph palette and Cursor legend in the figure below).
Each plot can be associated to either one of the two vertical axes by right-clicking on the plot icon and navigating down to the Y Scale menu:
and selecting the relevant axis. Plots that "don't look right" might just be associated with the wrong Y axis.
1. Average lifetime from phasor ratio analysis (Phasor Graph Panel)
Conversion from phasor to lifetime is not trivial, unless the phasor is located on the universal circle or is known to be comprised of a mixture of species with known lifetimes.
The convention used by AlliGator is to convert the phasor ratio into an average lifetime based on user-provided lifetimes for phasor reference 1 and 2.
To provide a phasor reference lifetime, simply type in a value in the Calibration Lifetime control (on the Phasor Graph panel) and right-click it to select either of the two following menu items:
Once this is done, any computation of a phasor ratio in the Phasor Ratio Analysis panel will add a point to the Lifetime Graph in the Lifetime Analysis panel, associated with the same abscissa as that used for the phasor ratio.
2. Phase lifetime (Phasor Graph Panel)
Another possible use of this representation is described in the Time Series Analysis manual page.
Note that, as for the Phasor Ratio Graph, it is possible to create a new empty plot to store new computed lifetimes in different plots.
Keep in mind that whenever a phasor ratio plot is created in the Phasor Ratio Graph, a plot is also created in the Lifetime Graph, and reciprocally.
3. Decay fit parameters (Fluorescence Decay Panel)
As discussed in the Fluorescence Decay Fitting manual page, it is possible to have the different parameters of a series of fits (whether independent or global) output as separate plots (one plot per selected parameter).
4. Decay shifts (Fluorescence Decay Panel)
Finally, the Plot Decay Shifts right-click menu item allows the set of stored decay shifts to be plotted (see the Fluorescence Decay manual page)
5. Building an Histogram of a Plot
The Plot Histogram right-click menu item allows building an histogram (with user-provided bin size) of the selected plot (as usual in Graphs, a plot can be selected either by right-clicking on its name in the Graph Legend, or directly on or close to the plot within the plot region). This opens the Histogram Window (described in the corresponding page of the manual), where a number of basic analyses can be carried out.
Page updated
Report abuse