Supplementary Figure 5: Correcting for differences in the excitation intensity in TIRF microscopy.
From: Precision and accuracy of single-molecule FRET measurements—a multi-laboratory benchmark study
Accounting for the differences in the excitation intensity profiles of the green and red laser across the field of view. The individual excitation profiles are determined as the mean image of a stack of images recorded while moving across a dense layer of dyes. In contrast to the uncorrected case (“before”), a position specific normalization creates narrower and more symmetric SE-populations (“after”). The standard corrections described in the main text are performed subsequently.
