Compressive three-dimensional super-resolution microscopy with speckle-saturated fluorescence excitation.
M PascucciS GanesanA TripathiO KatzValentina EmilianiM GuillonPublished in: Nature communications (2019)
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.
Keyphrases
- single molecule
- high resolution
- energy transfer
- high speed
- high density
- mass spectrometry
- magnetic resonance
- high throughput
- magnetic resonance imaging
- computed tomography
- endothelial cells
- cell cycle arrest
- cell proliferation
- quantum dots
- cell death
- label free
- photodynamic therapy
- contrast enhanced
- electron microscopy