Fast Inertia-Free Volumetric Light-Sheet Microscope

Abstract

Fast noninvasive three-dimensional (3D) imaging is crucial for quantitatively studying highly dynamic events ranging from flow cytometry to developmental biology. Light-sheet microscopy has emerged as the tool-of-choice for 3D characterization of rapidly evolving systems. However, to obtain a 3D image, either the sample or parts of the microscope are moved, limiting the acquisition speed. Here, we propose a novel inertia-free light-sheet-based scheme for volumetric imaging at high temporal resolution. Our approach comprises a novel combination of an acousto-optic scanner to produce tailored illumination and an acoustic-optofluidic lens, placed in the detection path to provide extended depth-of-field. Such a combination enables unsurpassed 3D imaging speeds up to 200 volumes per second. The volumetric imaging speed is limited only by the acquisition frame rate of the camera and the desired signal-to-noise ratio. Our volumetric microscope allows for invariant acquisition along the detection axis, avoiding extensive processing or complex deconvolution methods to restore image quality.