Open AccessWavefront engineered light needle microscopy for axially resolved rapid volumetric imaging
Author(s) -
Yuichi Kozawa,
Tomoya Nakamura,
Yuuki Uesugi,
Shunichi Sato
Publication year - 2022
Publication title -
biomedical optics express
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.449329
Subject(s) - raster scan , optics , microscopy , wavefront , materials science , laser scanning , optical sectioning , micrometer , light sheet fluorescence microscopy , image resolution , raster graphics , laser , scanning confocal electron microscopy , computer science , computer vision , physics
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.