Uniform and scalable light-sheets generated by extended focusing

Abstract

Light-sheet fluorescence microscopy (LSFM) affords highly parallelized 3D imaging with optical sectioning capability and minimal light exposure. However, using Gaussian beams for light-sheet generation results in a trade-off between beam waist thickness and the area over which the beam can approximate a light-sheet. Here, we present a novel form of LSFM that uses incoherent extended focusing to produce divergence free light-sheets with near diffraction-limited resolution and uniform intensity distribution along the propagation direction. We demonstrate the imaging performance of the new technique by volumetric imaging of beads, collagen fibers, and melanoma cancer cells with sub-cellular resolution.