Sample drift estimation method based on speckle patterns formed by backscattered laser light
Volodymyr Nechyporuk-Zloy
10%+ Growth Driver | Manager | Microscopist | DL Imaging
Sample drift estimation method based on speckle patterns formed by backscattered laser light
Shih-Ya Chen, Rainer Heintzmann, and Christoph Cremer
https://doi.org/10.1364/BOE.10.006462
?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.