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bioRxiv · 10.1101/2025.08.14.670427

Efficient Double Helix Detection with Steerable Filters

Barentine, A. E. S.·Balaji, A.·Moerner, W. E.
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Abstract

We present an efficient detection scheme for localization of Double Helix point-spread functions for 3D single-molecule localization microscopy or tracking. Using steerable filters, we extract both 2D position and lobe orientation (axial position) estimates using just 7 convolutions, orders of magnitude less than used in deep-learning-based approaches. We pair this detection with a fitter and implement both as a plug-in for the open source PYthon Microscopy Environment (PYME), which features percentile-based background subtraction, signal-to-noise-based detection thresholding, and performant parallel analysis. Our complete localization analysis pipeline achieves state-of-the-art performance with minimal user input.

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Barentine, A. E. S., Balaji, A., Moerner, W. E.. 2025-08-20. Efficient Double Helix Detection with Steerable Filters. https://doi.org/10.1101/2025.08.14.670427

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