Multi-focus light-field microscopy for high-speed large-volume imaging

Abstract

High-speed visualization of three-dimensional (3D) processes across a large field of view with cellular resolution is essential for understanding living systems. Light-field microscopy (LFM) has emerged as a powerful tool for fast volumetric imaging. However, one inherent limitation of LFM is that the achievable lateral resolution degrades rapidly with the increase of the distance from the focal plane, which hinders the applications in observing thick samples. Here, we propose Spherical-Aberration-assisted scanning LFM (SAsLFM), a hardware-modification-free method that modulates the phase-space point-spread-functions (PSFs) to extend the effective high-resolution range along the z-axis by ~ 3 times. By transferring the foci to different depths, we take full advantage of the redundant light-field data to preserve finer details over an extended depth range and reduce artifacts near the original focal plane. Experiments on a USAF-resolution chart and zebrafish vasculatures were conducted to verify the effectiveness of the method. We further investigated the capability of SAsLFM in dynamic samples by imaging large-scale calcium transients in the mouse brain, tracking freely-moving jellyfish, and recording the development of Drosophila embryos. In addition, combined with deep-learning approaches, we accelerated the three-dimensional reconstruction of SAsLFM by three orders of magnitude. Our method is compatible with various phase-space imaging techniques without increasing system complexity and can facilitate high-speed large-scale volumetric imaging in thick samples.

Virtual-scanning light-field microscopy for robust snapshot high-resolution volumetric imaging

“…The use of objective lenses with a lower numerical aperture would improve axial range at the cost of spatial resolution. Improvement of the angular resolution 20 or phase modulation at the pupil plane 51 is also anticipated to increase the axial coverage. As a one-photon fluorescence microscopy technique, the imaging penetration capability of VsLFM is fundamentally limited by tissue scattering and background fluorescence, which can be enhanced with other scattering removal methods 27 .…”

How I Met Your V2X Sensor Data: Analysis of Projection-Based Light Field Visualization for Vehicle-to-Everything Communication Protocols and Use Cases

“…Recent works on light field technology cover the topics of angular enhancement [ 100 , 101 , 102 ] (which aims to achieve super-resolution), image enhancement [ 103 , 104 , 105 ] (which is also often named super-resolution, and thus, the terminology of the other one is becoming “angular super-resolution”), saliency detection [ 106 , 107 , 108 ], light field rendering [ 109 , 110 , 111 ] and reconstruction [ 112 , 113 , 114 ], microscopy [ 115 , 116 , 117 ], camera animation [ 118 ], video streaming [ 119 ], objective quality assessment [ 120 , 121 ], perceived quality [ 94 , 122 ], and many more. Every single experiment that studies the projection-based light field visualization with the involvement of human individuals is summarized in a recent work [ 123 ], along with a thorough analysis of future research efforts, and an up-to-date survey [ 124 ] investigates the associated methods.…”

Aberration modeling in deep learning for volumetric reconstruction of light-field microscopy

“…Some works even model the aberration in deep learning networks [23][24][25] to well ease its influence and improve the imaging quality. However, the aberration problem is rarely explored in the field of LFM 5,26 . Wu et.al 5 combinedly utilize a scanning LFM system and iterative digital adaptive optics (DAO) to realize subcellular resolution and long-term observation of 3D dynamics.…”

“…Wu et.al 5 combinedly utilize a scanning LFM system and iterative digital adaptive optics (DAO) to realize subcellular resolution and long-term observation of 3D dynamics. In addition, Zhang et.al 26 propose introducing spherical aberration to expand the depth of field (DOF) of scanning LFM and reduce artifacts near the original focal plane. But to the best of our knowledge, no research has been proposed to achieve the snapshot 3D observation of live dynamic samples at micrometer resolution under the condition of heavy spherical aberration.…”