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bioRxiv · 10.1101/2023.01.26.525798

Extracting nanoscale morphology from localizations

Marin, Z.·Fuentes, L. A.·Bewersdorf, J.·Baddeley, D.
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Abstract

Membrane surface reconstruction at the nanometer scale is required for understanding mechanisms of subcellular shape change. This historically has been the domain of electron microscopy, but extraction of surfaces from specific labels is a difficult task in this imaging modality. Existing methods for extracting surfaces from fluorescence microscopy have poor resolution or require high-quality super-resolution data that is manually cleaned and curated. Here we present NanoWrap, a new method for extracting surfaces from generalized single-molecule localization microscopy (SMLM) data. This makes it possible to study the shape of specifically-labelled membraneous structures inside of cells. We validate NanoWrap using simulations and demonstrate its reconstruction capabilities on SMLM data of the endoplasmic reticulum and mitochondria. NanoWrap is implemented in the open-source Python Microscopy Environment. SIGNIFICANCEWe introduce a novel tool for reconstruction of subcellular membrane surfaces from single-molecule localization microscopy data and use it to visualize and quantify local shape and membrane-membrane interactions. We benchmark its performance on simulated data and demonstrate its fidelity to experimental data.

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Marin, Z., Fuentes, L. A., Bewersdorf, J., Baddeley, D.. 2023-01-27. Extracting nanoscale morphology from localizations. https://doi.org/10.1101/2023.01.26.525798

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