Particle Localization Using Local Gradients and Its Application to Nanometer Stabilization of a Microscope

Year: 2023

Authors: Kashchuk AV., Perederiy O., Caldini C., Gardini L., Pavone FS., Negriyko AM., Capitanio M.

Autors Affiliation: Inst Phys NASU, UA-03680 Kiev, Ukraine; LENS, European Lab Nonlinear Spect, I-50019 Sesto Fiorentino, Italy; CNR, Natl Inst Opt, I-50125 Florence, Italy; Univ Florence, Dept Phys & Astron, I-50019 Sesto Fiorentino, Italy.

Abstract: Particle localization plays a fundamental role in advanced biological techniques such as single-molecule tracking, superresolution microscopy, and manipulation by optical and magnetic tweezers. Such techniques require fast and accurate particle localization algorithms as well as nanometerscale stability of the microscope. Here, we present a universal method for three-dimensional localization of single labeled and unlabeled particles based on local gradient calculation of particle images. The method outperforms state-of-the-art localization techniques in high-noise conditions, and it is capable of 3D nanometer accuracy localization of nano-and microparticles with sub-millisecond calculation time. By localizing a fixed particle as fiducial mark and running a feedback loop, we demonstrate its applicability for active drift correction in sensitive nanomechanical measurements such as optical trapping and superresolution imaging. A multiplatform open software package comprising a set of tools for local gradient calculation in brightfield, darkfield, and fluorescence microscopy is shared for ready use by the scientific community.

Journal/Review: ACS NANO

Volume: 17 (2)      Pages from: 1344  to: 1354

More Information: This work was supported by the European Union’s Horizon 2020 research and innovation program under grant agreement no. 871124 Laserlab-Europe. A.V.K . was supported by the Human Frontier Science Program Cross-Disciplinary Fellowship LT008/2020-C. O.P. and A.M.N. were supported by the project 1.4. B/185 National Academy of Sciences of Ukraine. A.V.K., O.P., and A.M.N. thank the Armed Forces of Ukraine for providing security and making this work possible.
KeyWords: particle tracking; microscope stabilization; 3D localization; radial symmetry; local gradients; fluorescence microscopy
DOI: 10.1021/acsnano.2c09787

Citations: 3
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