TY - JOUR T1 - 3D Interferometric Lattice Light-Sheet Imaging JF - bioRxiv DO - 10.1101/2020.08.27.266999 SP - 2020.08.27.266999 AU - Bin Cao AU - Guanshi Wang AU - Jieru Li AU - Alexandros Pertsinidis Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/08/28/2020.08.27.266999.abstract N2 - Understanding cellular structure and function requires live-cell imaging with high spatio-temporal resolution and high detection sensitivity. Direct visualization of molecular processes using single-molecule/super-resolution techniques has thus been transformative. However, extracting the highest-resolution 4D information possible from weak and dynamic fluorescence signals in live cells remains challenging. For example, some of the highest spatial resolution methods, e.g. interferometric (4Pi) approaches1-6 can be slow, require high peak excitation intensities that accelerate photobleaching or suffer from increased out-of-focus background. Selective-plane illumination (SPIM)7-12 reduces background, but most implementations typically feature modest spatial, especially axial, resolution. Here we develop 3D interferometric lattice light-sheet (3D-iLLS) imaging, a technique that overcomes many of these limitations. 3D-iLLS provides, by virtue of SPIM, low light levels and photobleaching, while providing increased background suppression and significantly improved volumetric imaging/sectioning capabilities through 4Pi interferometry. We demonstrate 3D-iLLS with axial resolution and single-particle localization precision down to <100nm (FWHM) and <10nm (1σ) respectively. 3D-iLLS paves the way for a fuller elucidation of sub-cellular phenomena by enhanced 4D resolution and SNR live imaging.Competing Interest StatementMSKCC has filed a patent application on modulation interferometric imaging systems and methods. ER -