%0 Journal Article %A Clément Cabriel %A Nicolas Bourg %A Pierre Jouchet %A Guillaume Dupuis %A Christophe Leterrier %A Aurélie Baron %A Marie-Ange Badet-Denisot %A Boris Vauzeilles %A Emmanuel Fort %A Sandrine Lévêque-Fort %T Combining 3D single molecule localization strategies for reproducible bioimaging %D 2018 %R 10.1101/385799 %J bioRxiv %P 385799 %X We developed a 3D localization-based super-resolution technique providing an almost isotropic 3D resolution 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. 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. %U https://www.biorxiv.org/content/biorxiv/early/2018/08/06/385799.full.pdf