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

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Applied Physics Letters. 2023 Jul 13;123(2). doi: 10.1063/5.0137834

Hinterer F, Schneider MC, Hubmer S, López-Martínez M, Ramlau R, Schütz GJ

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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. Particularly recording at cryogenic temperatures dramatically reduces photobleaching and would, hence, in principle, allow the user to massively increase the illumination time to several seconds. The downside of long illuminations, however, would be image blur due to inevitable jitter or drift occurring during the illuminations, which deteriorates the localization precision. In this paper, we theoretically demonstrate that a parallel recording of the fiducial marker beads together with a fitting approach accounting for the full drift trajectory allows for largely eliminating drift effects for drift magnitudes of several hundred nanometers per frame. We showcase the method for linear and diffusional drift as well as oscillations, assuming fixed dipole orientations during each illumination.

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

Abstract

Janelia Authors