Combining 3D single molecule localization strategies for reproducible bioimaging

Clément Cabriel (), Nicolas Bourg, Pierre Jouchet, Guillaume Dupuis, Christophe Leterrier, Aurélie Baron, Marie-Ange Badet-Denisot, Boris Vauzeilles, Emmanuel Fort and Sandrine Lévêque-Fort ()
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Clément Cabriel: CNRS, Univ. Paris-Sud, Université Paris-Saclay
Nicolas Bourg: CNRS, Univ. Paris-Sud, Université Paris-Saclay
Pierre Jouchet: CNRS, Univ. Paris-Sud, Université Paris-Saclay
Guillaume Dupuis: Univ. Paris-Sud, Université Paris-Saclay, CNRS, Fédération LUMAT
Christophe Leterrier: Aix-Marseille Université, CNRS, INP, NeuroCyto
Aurélie Baron: Institut de Chimie des Substances Naturelles du CNRS
Marie-Ange Badet-Denisot: Institut de Chimie des Substances Naturelles du CNRS
Boris Vauzeilles: Institut de Chimie des Substances Naturelles du CNRS
Emmanuel Fort: ESPCI Paris, PSL University, CNRS
Sandrine Lévêque-Fort: CNRS, Univ. Paris-Sud, Université Paris-Saclay

Nature Communications, 2019, vol. 10, issue 1, 1-10

Abstract: 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.

Date: 2019
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DOI: 10.1038/s41467-019-09901-8

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