High-speed 4D fluorescence light field tomography of whole freely moving organisms.

IF 8.5 1区 物理与天体物理 Q1 OPTICS
Optica Pub Date : 2025-05-20 Epub Date: 2025-05-05 DOI:10.1364/optica.549707
Kevin C Zhou, Clare Cook, Archan Chakraborty, Jennifer Bagwell, Joakim Jönsson, Kyung Chul Lee, Xi Yang, Shiqi Xu, Ramana Balla, Kushal Kolar, Caitlin Lewis, Mark Harfouche, Donald T Fox, Michel Bagnat, Roarke Horstmeyer
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引用次数: 0

Abstract

Volumetric fluorescence imaging techniques, such as confocal, multiphoton, light sheet, and light field microscopy, have become indispensable tools across a wide range of cellular, developmental, and neurobiological applications. However, it is difficult to scale such techniques to the large 3D fields of view (FOV), volume rates, and synchronicity requirements for high-resolution 4D imaging of freely behaving organisms. Here, we present reflective Fourier light field computed tomography (ReFLeCT), a high-speed volumetric fluorescence computational imaging technique. ReFLeCT synchronously captures entire tomograms of multiple unrestrained, unanesthetized model organisms across multi-millimeter 3D FOVs at 120 volumes per second. In particular, we applied ReFLeCT to reconstruct 4D videos of fluorescently labeled zebrafish and Drosophila larvae, enabling us to study their heartbeat, fin and tail motion, gaze, jaw motion, and muscle contractions with nearly isotropic 3D resolution while they are freely moving. To our knowledge, as a novel approach for snapshot tomographic capture, ReFLeCT is a major advance toward bridging the gap between current volumetric fluorescence microscopy techniques and macroscopic behavioral imaging.

整个自由活动生物体的高速四维荧光场断层扫描。
体积荧光成像技术,如共聚焦、多光子、光片和光场显微镜,已经成为广泛的细胞、发育和神经生物学应用中不可或缺的工具。然而,很难将这种技术扩展到大3D视场(FOV),体积率和高分辨率四维成像要求的同步性。在这里,我们提出反射傅里叶光场计算机断层扫描(ReFLeCT),一种高速体积荧光计算成像技术。ReFLeCT以每秒120个体积的速度同步捕获多个不受约束、未麻醉的模型生物在多毫米3D fov上的整个层析成像。特别是,我们应用ReFLeCT重建了荧光标记的斑马鱼和果蝇幼虫的4D视频,使我们能够以几乎各向同性的3D分辨率研究它们自由运动时的心跳,鳍和尾巴运动,凝视,下巴运动和肌肉收缩。据我们所知,作为一种快照层析成像的新方法,ReFLeCT是弥合当前体积荧光显微镜技术和宏观行为成像技术之间差距的重大进步。
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来源期刊
Optica
Optica OPTICS-
CiteScore
19.70
自引率
2.90%
发文量
191
审稿时长
2 months
期刊介绍: Optica is an open access, online-only journal published monthly by Optica Publishing Group. It is dedicated to the rapid dissemination of high-impact peer-reviewed research in the field of optics and photonics. The journal provides a forum for theoretical or experimental, fundamental or applied research to be swiftly accessed by the international community. Optica is abstracted and indexed in Chemical Abstracts Service, Current Contents/Physical, Chemical & Earth Sciences, and Science Citation Index Expanded.
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