Phasor based single-molecule localization microscopy in 3D (pSMLM-3D): An algorithm for MHz localization rates using standard CPUs
Koen J.A. Martens (Wageningen University & Research)
Arjen N. Bader (Wageningen University & Research)
Sander Baas (Wageningen University & Research)
B Rieger (TU Delft - ImPhys/Quantitative Imaging)
Johannes Hohlbein (Wageningen University & Research)
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Abstract
We present a fast and model-free 2D and 3D single-molecule localization algorithm that allows more than 3 × 106 localizations per second to be calculated on a standard multi-core central processing unit with localization accuracies in line with the most accurate algorithms currently available. Our algorithm converts the region of interest around a point spread function to two phase vectors (phasors) by calculating the first Fourier coefficients in both the x- and y-direction. The angles of these phasors are used to localize the center of the single fluorescent emitter, and the ratio of the magnitudes of the two phasors is a measure for astigmatism, which can be used to obtain depth information (z-direction). Our approach can be used both as a stand-alone algorithm for maximizing localization speed and as a first estimator for more time consuming iterative algorithms.
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