Phase-modulated multi-foci microscopy for rapid 3D imaging

3D imaging technology is pivotal in monitoring the functional dynamics and morphological alterations in living cells and tissues.However,conventional volumetric imaging associated with mechanical z-scanning encounters challenges in limited 3D imaging speed with inertial artifact.Here,we present a unique phase-modulated multifoci microscopy (PM^(3)) technique to achieve snapshot 3D imaging with the advantages of extended imaging depths and adjustable imaging intervals between each focus in a rapid fashion.To accomplish the tasks,we utilize a spatial light modulator (SLM) to encode the phases of the scattered or fluorescence light emanating from a volumetric sample and then project the multiple-depth images of the sample onto a single charge-coupled device camera for rapid 3D imaging.We demonstrate that the PM^(3)technique provides~55-fold improvement in imaging depth in polystyrene beads phantom compared to the depth of field of the objective lens used.PM^(3)also enables the real-time monitoring of Brownian motion of fluorescent beads in water at a 15 Hz volume rate.By precisely manipulating the phase of scattered light on the SLM,PM^(3)can pinpoint the specific depth information in living zebrafish and rapidly observe the 3D dynamic processes of blood flow in the zebrafish trunk.This work shows that the PM^(3)technique developed is robust and versatile for fast 3D dynamic imaging in biological and biomedical systems.