
Sample drift estimation method based on speckle patterns formed by backscattered laser light
Author(s) -
ShihYa Chen,
Rainer Heintzmann,
Christoph Cremer
Publication year - 2019
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.10.006462
Subject(s) - speckle pattern , optics , microscope , fiducial marker , laser , stack (abstract data type) , microscopy , sample (material) , image resolution , nanometre , resolution (logic) , materials science , physics , computer science , artificial intelligence , thermodynamics , programming language
Single molecule localization microscopy (SMLM) has been established to acquire images with unprecedented resolution down to several nanometers. A typical time scale for image acquisition is several minutes to hours. Yet it is difficult to avoid completely sample drift for long time measurements. To estimate drift, we present a method based on the evaluation of speckle patterns formed by backscattered laser light from the cells using a single molecule localization microscope setup. A z-stack of unique speckle patterns is recorded prior to the measurements as a three-dimensional position reference. During the experiment, images of scattered laser light were acquired, and correlated individually with each of the images of the speckle reference stack to estimate x, y and z drift. Our method shows highly comparable results with a fiducial marker approach, achieving a precision of several nanometers. This method allows for high precision three dimensional drift correction of microscope systems without any additional sample preparation.