We determine the optimal parameters (scan velocities) for measuring the luminescence lifetime on the microsecond scale using the recently introduced method based on scanning the excitation beam across the sample. Using simulations, we evaluate the standard deviation and bias of the luminescence decay rate determined by scanning with two different velocities. The analysis is performed for Poisson- and normal-distributed signals, representing different types of detection techniques. We also show that a weak uncorrected background induces a bias in the obtained decay rate, and take this effect into account when choosing optimal measurement parameters. For comparison, the analysis is additionally performed for two conventional gating schemes for lifetime measurement. The variable-velocity scanning method is found to be more robust to the effect of the background signal than the gating schemes.
- confocal laser scanning microscopy
- error analysis
- lifetime imaging
ASJC Scopus subject areas
- Medical Laboratory Technology