2021/09/02 by Masullo, Luciano A., Lopez, Lucía F., Stefani, Fernando D.
#Biological Physics (physics.bio-ph) #FOS: Physical sciences #Optics (physics.optics)
paper · doi:10.48550/arxiv.2109.01234
Localization of single fluorescent molecules is key for physicochemical and biophysical measurements such as single-molecule tracking and super-resolution imaging by single-molecule localization microscopy (SMLM). Recently a series of methods have been developed in which the localization precision is enhanced by interrogating the molecular position with a sequence of spatially modulated patterns of light. Among them, the MINFLUX technique outstands for achieving a ~10-fold improvement compared to wide-field camera-based single-molecule localization, reaching ~1-2 nm localization precision at moderate photon counts. Here, we present a common mathematical framework for this type of measurement that allows a fair comparison between reported methods and facilitates the design and evaluation of new methods. With it, we benchmark all reported methods for single-molecule localization using sequential structured illumination, including long-established methods such as orbital tracking, along with two new proposed methods: orbital tracking and raster scanning with a minimum of intensity.