Compressive three-dimensional super-resolution microscopy with speckle-saturated fluorescence excitation

M. Pascucci, S. Ganesan, A. Tripathi, O. Katz, V. Emiliani and M. Guillon ()
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M. Pascucci: University Paris Descartes
S. Ganesan: University Paris Descartes
A. Tripathi: The Hebrew University of Jerusalem
O. Katz: The Hebrew University of Jerusalem
V. Emiliani: University Paris Descartes
M. Guillon: University Paris Descartes

Nature Communications, 2019, vol. 10, issue 1, 1-8

Abstract: Abstract Nonlinear structured illumination microscopy (nSIM) is an effective approach for super-resolution wide-field fluorescence microscopy with a theoretically unlimited resolution. In nSIM, carefully designed, highly-contrasted illumination patterns are combined with the saturation of an optical transition to enable sub-diffraction imaging. While the technique proved useful for two-dimensional imaging, extending it to three-dimensions is challenging due to the fading of organic fluorophores under intense cycling conditions. Here, we present a compressed sensing approach that allows 3D sub-diffraction nSIM of cultured cells by saturating fluorescence excitation. Exploiting the natural orthogonality of speckles at different axial planes, 3D probing of the sample is achieved by a single two-dimensional scan. Fluorescence contrast under saturated excitation is ensured by the inherent high density of intensity minima associated with optical vortices in polarized speckle patterns. Compressed speckle microscopy is thus a simple approach that enables 3D super-resolved nSIM imaging with potentially considerably reduced acquisition time and photobleaching.

Date: 2019
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DOI: 10.1038/s41467-019-09297-5

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