Abstract

Super-resolution (SR) microscopy has dramatically enhanced our understanding of biological processes. However, scattering media in thick specimens severely limits the spatial resolution, often rendering the images unclear or indistinguishable. Additionally, live-cell imaging faces challenges in achieving high temporal resolution for fast-moving subcellular structures.

Here, we present the principles of a synthetic wave microscopy (SWM) to extract three-dimensional information from thick unlabeled specimens, where photobleaching and phototoxicity are avoided. SWM exploits multiple-wave interferometry to reveal the specimen’s phase information in the area of interest, which is not affected by the scattering media in the optical path. SWM achieves ~0.42 λ/NA resolution at an imaging speed of up to 106 pixels/s. SWM proves better temporal resolution and sensitivity than the most conventional microscopes currently available while maintaining exceptional SR and anti-scattering capabilities.

Penetrating through the scattering media is challenging for conventional imaging techniques. Remarkably, SWM retains its efficacy even in conditions of low signal-to-noise ratios. It facilitates the visualization of dynamic subcellular structures in live cells, encompassing tubular endoplasmic reticulum (ER), lipid droplets, mitochondria, and lysosomes.