Robust and bias-free localization of individual fixed dipole emitters achieving the Cramér Rao bound for applications in cryo-single molecule localization microscopy

Robust and bias-free localization of individual fixed dipole emitters achieving the Cramér Rao bound for applications in cryo-single molecule localization microscopy

Compared to room temperature experiments, SMLM performed under cryogenic temperature achieves higher photon yields and, hence, higher localization precision. However, to fully exploit the resolution it is crucial to account for the anisotropic emission characteristics of fluorescence dipole emitters with fixed orientation. We show here that astigmatic imaging in combination with information about the dipole orientation allows to extract the position of the dipole emitters without localization bias and down to a precision of 1 nm, thereby reaching the corresponding Cramér Rao bound.

Authors:

Fabian Hinterer¹, Magdalena C Schneider², Simon Hubmer³, Montserrat López-Martinez², Philipp Zelger⁴, Alexander Jesacher⁴, Ronny Ramlau^1 3, Gerhard J Schütz²

Reference:

PLOS ONE, Published: February 4, 2022 https://doi.org/10.1371/journal.pone.0263500