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
{"title":"High-speed 4D fluorescence light field tomography of whole freely moving organisms.","authors":"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","doi":"10.1364/optica.549707","DOIUrl":null,"url":null,"abstract":"<p><p>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 <i>Drosophila</i> 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.</p>","PeriodicalId":19515,"journal":{"name":"Optica","volume":"12 5","pages":"674-684"},"PeriodicalIF":8.5000,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12302686/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Optica","FirstCategoryId":"101","ListUrlMain":"https://doi.org/10.1364/optica.549707","RegionNum":1,"RegionCategory":"物理与天体物理","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/5/5 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"OPTICS","Score":null,"Total":0}
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.
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.