In standard SMLM methods, the photoswitching of single fluorescent molecules and the
data acquisition processes are independent, which leads to the detection of single
molecule blinking events on several consecutive frames. This mismatch results in several
data points with reduced localization precision, and it also increases the possibilities
of overlapping. Here we discuss how the synchronization of the fluorophores’ ON state
to the camera exposure time increases the average intensity of the captured point
spread functions and hence improves the localization precision. Simulations and theoretical
results show that such synchronization leads to fewer localizations with 15% higher
sum signal on average, while reducing the probability of overlaps by 10%.
