Temporal resolution

Physics

Spinning

Fluorescence microscope

Image resolution

Optics

Resolution (logic)

Microscopy

Deconvolution

Artificial intelligence

Composite material

Super-resolution microscopy

Materials science

Computer science

Microscope

Fluorescence

Nature Biotechnology

A main determinant of the spatial resolution of live-cell super-resolution (SR) microscopes is the maximum photon flux that can be collected. To further increase the effective resolution for a given photon flux, we take advantage of a priori knowledge about the sparsity and continuity of biological structures to develop a deconvolution algorithm that increases the resolution of SR microscopes nearly twofold. Our method, sparse structured illumination microscopy (Sparse-SIM), achieves ~60-nm resolution at a frame rate of up to 564 Hz, allowing it to resolve intricate structures, including small vesicular fusion pores, ring-shaped nuclear pores formed by nucleoporins and relative movements of inner and outer mitochondrial membranes in live cells. Sparse deconvolution can also be used to increase the three-dimensional resolution of spinning-disc confocal-based SIM, even at low signal-to-noise ratios, which allows four-color, three-dimensional live-cell SR imaging at ~90-nm resolution. Overall, sparse deconvolution will be useful to increase the spatiotemporal resolution of live-cell fluorescence microscopy. The resolution of fluorescence microscopy is increased by incorporating prior information into deconvolution algorithms.

Sparse deconvolution improves the resolution of live-cell super-resolution fluorescence microscopy