Localization of fixed dipoles at high precision by accounting for sample drift during illumination

TL;DR

This paper presents a novel method for high-precision localization of fixed dipoles in single molecule microscopy by modeling and correcting sample drift during long illumination periods, especially at cryogenic temperatures.

Contribution

It introduces a new fitting approach that accounts for the full drift trajectory, significantly improving localization accuracy during extended imaging sessions.

Findings

Drift correction method effectively reduces localization errors.

Longer illumination times at cryogenic temperatures enhance photon detection.

Sample drift can be largely mitigated with the proposed approach.

Abstract

Single molecule localization microscopy relies on the precise quantification of the position of single dye emitters in a sample. This precision is improved by the number of photons that can be detected from each molecule. It is therefore recommendable to increase illumination times for the recording process. Particularly recording at cryogenic temperatures dramatically reduces photobleaching and thereby allows a massive increase in illumination times to several seconds. As a downside, microscope instabilities may well introduce jitter during such long illuminations, deteriorating the localization precision. In this paper, we theoretically demonstrate that a parallel recording of fiducial marker beads together with a novel fitting approach accounting for the full drift trajectory allows for largely eliminating drift effects for drift magnitudes of several hundred nanometers per frame.