Embedded Files
It is possible to pre-process the data during or right after loading it. A bit counter-intuitively, these options can be found in Settings>>Fluorescence Decay>>Basic Analysis panel, as they are indeed relevant to decay (and phasor) analysis.
The operations that affect data are listed below, with a short description of their effect on the data.
1. Skipping Gates
Parameters that control which gates are retained from the original dataset are circled in red in the snapshot below:
Gate Step indicates whether all gates are kept (Gate Step = 1), or every other gate (Gate Step = 2), or every three gate, etc.
Gates to Skip specifies how many gates at the beginning (Start) and at the end (End) of the series are omitted when loading the file in memory. For instance, to keep all gates, both Start and End should be set to 0 (the default values). This option is useful to omit gates for which it is known that some recording artifact was present (low or high signal, for instance), or to "shift' the decay toward time 0 by dropping gates at the beginning of the recording corresponding to time windows before the laser pulse.
Note that changes to these parameters are only taken into account after reloading the current file (Ctrl+R).
2. Pile-up Correction
When pile-up is known to occur (as in SPAD arrays), it is generally easy (and necessary) to correct mathematically. Correction for pile-up due to 1-bit counters such as those found in Swiss SPAD 1 & 2 is currently implemented.To apply this correction, the Pile-up Correction checkbox in the Basic Analysis panel should be checked (see above). This reveals the Max Value parameter, which corresponds to the maximum number of recorded counts per pixel. For instance, for Swiss SPAD 2 (SS2) where 8-bit and 10-bit acquisition is possible, the Max Value parameter is respectively 255 and 255 x 4 = 1020, as the 10-bit image is in fact comprised of 4 8-bit images summed in software. If the number of acquired 1-bit frames is different, this new value should be used.
Note that changes to these parameters are only taken into account after reloading the current file (Ctrl+R).Â
3. Additional Operations
Additional decay pre-processing operations are described in the Fluorescence Decay page of the manual.
Page updated
Report abuse