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Journal Articles Nature Communications Year : 2019

Combining 3D single molecule localization strategies for reproducible bioimaging

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Abstract

Here, we present a 3D localization-based super-resolution technique providing a slowly varying localization precision over a 1 μm range with precisions down to 15 nm. The axial localization is performed through a combination of point spread function (PSF) shaping and supercritical angle fluorescence (SAF), which yields absolute axial information. Using a dual-view scheme, the axial detection is decoupled from the lateral detection and optimized independently to provide a weakly anisotropic 3D resolution over the imaging range. This method can be readily implemented on most homemade PSF shaping setups and provides drift-free, tilt-insensitive and achromatic results. Its insensitivity to these unavoidable experimental biases is especially adapted for multicolor 3D super-resolution microscopy, as we demonstrate by imaging cell cytoskeleton, living bacteria membranes and axon periodic submembrane scaffolds. We further illustrate the interest of the technique for biological multicolor imaging over a several-μm range by direct merging of multiple acquisitions at different depths.
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Dates and versions

hal-02116025 , version 1 (30-04-2019)

Licence

Attribution - CC BY 4.0

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Cite

Clément Cabriel, Nicolas Bourg, Pierre Jouchet, Guillaume Dupuis, Christophe Leterrier, et al.. Combining 3D single molecule localization strategies for reproducible bioimaging. Nature Communications, 2019, 10 (1), pp.1642. ⟨10.1038/s41467-019-09901-8⟩. ⟨hal-02116025⟩
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