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.
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.