Abstract
Increasing the acquisition speed of three-dimensional volumetric images is important—particularly in biological imaging—to unveil the structural dynamics and functionalities of specimens in detail. In conventional laser scanning fluorescence microscopy, volumetric images are constructed from optical sectioning images sequentially acquired by changing the observation plane, limiting the acquisition speed. Here, we present a novel method to realize volumetric imaging from two-dimensional raster scanning of a light needle spot without sectioning, even in the traditional framework of laser scanning microscopy. Information from multiple axial planes is simultaneously captured using wavefront engineering for fluorescence signals, allowing us to readily survey the entire depth range while maintaining spatial resolution. This technique is applied to real-time and video-rate three-dimensional tracking of micrometer-sized particles, as well as the prompt visualization of thick fixed biological specimens, offering substantially faster volumetric imaging.
Funder
Precursory Research for Embryonic Science and Technology
Japan Society for the Promotion of Science
Japan Agency for Medical Research and Development
Konica Minolta Science and Technology Foundation
Dynamic Alliance for Open Innovation Bridging Human, Environment and Materials
Subject
Atomic and Molecular Physics, and Optics,Biotechnology
Cited by
7 articles.
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