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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 new high-speed volumetric fluorescence computational imaging technique. ReFLeCT synchronously captures entire tomograms of multiple unrestrained, unanesthetized model organisms over 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. As a novel approach for snapshot tomographic capture, ReFLeCT is a major advance towards bridging the gap between current volumetric fluorescence microscopy techniques and macroscopic behavioral imaging.
This data repository contains the raw data for performing 4D reconstructions for four of the samples featured in the accompanying publication (link TBD). The code that processes this data can be found here: https://github.com/kevinczhou/ReFLeCT.... [Read More]
Total Size
5 files (44.6 GB)
Data Citation
Zhou, K., Cook, C., Chakraborty, A., Bagwell, J., Jönsson, J., Lee, K. C., Yang, X., Xu, S., Balla, R., Kolar, K., Lewis, C., Harfouche, M., Fox, D., Bagnat, M., & Horstmeyer, R. (2025). Data from: High-speed 4D fluorescence light field tomography of whole freely moving organisms. Duke Research Data Repository. https://doi.org/10.7924/r4zs30t4h
Zhou, K., Cook, C., Chakraborty, A., Bagwell, J., Jönsson, J., Lee, K. C., Yang, X., Xu, S., Balla, R., Kolar, K., Lewis, C., Harfouche, M., Fox, D., Bagnat, M., & Horstmeyer, R. (2025). High-speed 4D fluorescence light field tomography of whole freely moving organisms. Optica, https://doi.org/10.1364/OPTICA.549707
Zhou, K., Cook, C., Chakraborty, A., Bagwell, J., Jönsson, J., Lee, K. C., Yang, X., Xu, S., Balla, R., Kolar, K., Lewis, C., Harfouche, M., Fox, D., Bagnat, M., & Horstmeyer, R. (2024). High-speed 4D fluorescence light field tomography of whole freely moving organisms. bioRxiv preprint: https://www.biorxiv.org/content/10.1101/2024.09.16.609432v1
Research reported in this publication was supported by the Office of Research Infrastructure Programs (ORIP), Office Of The Director, National Institutes Of Health of the National Institutes Of Health and the National Institute Of Environmental Health Sciences (NIEHS) of the National Institutes of Health under Award Number R44OD024879, the National Cancer Institute (NCI) of the National Institutes of Health under Award Number R44CA250877, the National Institute of Biomedical Imaging and Bioengineering (NIBIB) of the National Institutes of Health under Award Number R43EB030979, the National Institute of General Medical Sciences of the National Institutes of Health under Award Numbers R01GM150666, R01GM118447 and R01GM140138, the National Science Foundation under Award Number 2036439, a Duke-Coulter Translational Partnership Grant, the American Heart Association under Award Number 23POST1013432. This work was also supported in part by the Schmidt Science Fellows, in partnership with the Rhodes Trust.
