Volumetric interferometric lattice light-sheet imaging

Live cell imaging with high spatiotemporal resolution and high detection sensitivity facilitates the study of the dynamics of cellular structure and function. However, extracting high-resolution 4D (3D space plus time) information from live cells remains challenging, because current methods are slow, require high peak excitation intensities or suffer from high out-of-focus background. Here we present 3D interferometric lattice light-sheet (3D-iLLS) imaging, a technique that requires low excitation light levels and provides high background suppression and substantially improved volumetric resolution by combining 4Pi interferometry with selective plane illumination. We demonstrate that 3D-iLLS has an axial resolution and single-particle localization precision of 100 nm (FWHM) and <10 nm (1 sigma), respectively. We illustrate the performance of 3D-iLLS in a range of systems: single messenger RNA molecules, nanoscale assemblies of transcription regulators in the nucleus, the microtubule cytoskeleton and mitochondria organelles. The enhanced 4D resolution and increased signal-to-noise ratio of 3D-iLLS will facilitate the analysis of biological processes at the sub-cellular level. New lattice light-sheet microscopy approach improves live cell imaging.

Automated summary

The 3D-iLLS imaging technique offers high background suppression and significantly improved volumetric resolution by using low excitation light levels and combining 4Pi interferometry with selective plane illumination. It demonstrates high axial resolution and single-particle localization precision, and shows strong performance in various systems, enhancing 4D resolution and signal-to-noise ratio for analyzing biological processes at the sub-cellular level.