Nanotime Analysis

The Nanotime Analysis page is comprised of two graphs:

• Fluorescence Decay Curves

• Fluorescence Decay Residuals

The former graph display three types of curves:

1. Instrument response functions (IRF)

2. Fluorescence lifetime histograms

3. Fits of fluorescence lifetime histograms

The latter graph  displays residuals (raw or normalized) of the previously mentioned fits. The residuals  are defined as:

where hi are the histogram values and fi the corresponding fit values.

The normalized residuals are defined as:

1. Fluorescence Decay Curves

1.1. Instrument Response Functions

IRFs are exported from the Nanotime Display page graphs as explained in the corresponding manual page. Their name indicate the type of IRF (Donor or Acceptor) selected by the user.

The plots are by default associated with the right-hand side scale, which is preset to show curves in Log scale.

Having two different scales for IRFs and experimetnal decays is useful to superimpose them, because of their very different range (IRFs are base-line corrected and normalized, while experimental histograms are not).

1.2. Nanotime Histograms

Experimental nanotime histograms are exported from the Nanotime Display page graphs as explained in the corresponding manual page.

The plots are by default associated with the left-hand side scale. Their name indicate the selection performed by the user when exporting the histogram.

1.3. Fits

Nanotime histogram fits are plotted in that same graph, as soon as a Nanotime Histogram Analysis is started .

The fit plots are by default associated with the left-hand side scale. Their name indicate which nanotime histogram decay they correspond to,

2. Nanotime Histogram Analysis

Nanotime histogram analysis requires at least one IRF and one compatible nanotime histogram to be present in the Fluorescence Decay Curves graph.

Right-clicking on the nanotime histogram of interest gives access to the following shortcut menu:

Selecting Fit Nanotime Histogram (1 Plot) will open the Nanotime Histogram Analysis window, described in the corresponding manual page.

Similarly, if there are two nanotime histograms (with their respective IRF(s)), the Fit Nanotime Histogram (2 Plots) shortcut will allow the user to successively select two plots to be fitted globally with the help of the Nanotime Histogram Analysis window, as described in the corresponding manual page.

3. Fluorescence Decay Residuals

The Fluorescence Decay Residuals graph displays the residuals of the fitted curves, according to the Normalized Residuals checkbox option. If the value of the checkbox is changed, the residual curve is only updated if the  Nanotime Histogram Analysis window computes a new curve. In other words, this checkbox has no effect on the plotted curves once the fit procedure is completed.