Single-molecule localization microscopy is a super-resolution imaging technique that relies on the spatial and temporal separation of blinking fluorescent emitters. Individual localization is possible for these blinking events with a precision significantly smaller than the classical diffraction limit.
This sub-diffraction localization precision depends on the number of photons emitted per molecule and sensor noise. We can estimate these parameters from the raw images. Alternatively, a rendered image of the localizations can provide an estimation of the resolution.
The rendering of localization datasets (single-molecule localization microscopy) can influence the resolution estimation based on decorrelation analysis. A modified histogram rendering, termed bilinear histogram, circumvents the biases introduced by Gaussian or standard histogram rendering. Researchers propose a parameter-free processing pipeline. They show that the resolution estimation becomes a function of the localization density and the localization precision on both simulated and state-of-the-art experimental datasets.