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

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 (3D) is challenging due to the fading/fatigue of organic fluorophores under intense cycling conditions. Here, we present a compressed sensing approach that allows for the first time 3D sub-diffraction nSIM of cultured cells by saturating fluorescence excitation. Exploiting the natural orthogonality of transverse speckle illumination 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.les fast 3D super-resolved imaging with considerably minimized photo-bleaching.

References in corpus (6)

• Exploiting speckle correlations to improve the resolution of wide-field fluorescence microscopy
• Customizing Speckle Intensity Statistics
• Structured illumination microscopy with unknown patterns and a statistical prior
• Joint reconstruction strategy for structured illumination microscopy with unknown illuminations
• Photoacoustic imaging beyond the acoustic diffraction-limit with dynamic speckle illumination and sparse joint support recovery
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