Single-Molecule Localization Microscopy Data Rendering

Single-molecule localization microscopy is a super-resolution imaging technique that relies on blinking fluorescent emitters’ spatial and temporal separation. 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 estimate 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.

In 2019, they proposed a novel method for estimating resolution on a single image using decorrelation analysis. A partial phase autocorrelation for a series of filtered images determines the highest spatial frequency with a sufficiently high signal-to-noise ratio. The method has been tested across imaging modalities for over a year using open-source software.

They’ve gotten mostly positive feedback from experts ranging from two-photon microscopy to structured illumination imaging. Initially, they presented decorrelation analysis at the single-molecule localization microscopy symposium (SMLMS), hoping for feedback from this image-processing-heavy community.

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