Theoretical Comparison of Real-Time Single Particle Tracking Techniques

نویسندگان

چکیده

One of the main challenges in studying single biomolecules a native or near-native environment is their constant diffusive motion. A method capable keeping biomolecule detection volume without disturbing its surrounding real-time particle tracking (SPT). There are number SPT methods use, and comparisons literature usually made based on experimental results, which, due to variations setups specimens, limited use. In this study, we used statistical calculations dynamic simulations compare theoretical performance different localization methods, namely orbital method, Knight's Tour (grid scan) MINFLUX, context both fluorescence-based interferometric scattering (iSCAT) approaches. While able track fastest diffusion, MINFLUX achieve greatest precision, albeit only for slowly diffusing particles. For comparing iSCAT fluorescence, considered biological examples, including intrinsically fluorescent particles with label. The relative success vs fluorescence strongly dependent size, photophysical properties fluorophore, fluorophore density. This work should serve as guideline choosing suitable implementation sample interest. analysis can easily be extended other interest that have not considered.

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ژورنال

عنوان ژورنال: Biophysical Journal

سال: 2021

ISSN: ['0006-3495', '1542-0086']

DOI: https://doi.org/10.1016/j.bpj.2020.11.1284