title: Mesoscopic in vivo 3-D tracking of sparse cell populations using angular multiplexed optical projection tomography.
abstract: We describe an angular multiplexed imaging technique for 3-D in vivo cell tracking of sparse cell distributions and optical projection tomography (OPT) with superior time-lapse resolution and a significantly reduced light dose compared to volumetric time-lapse techniques. We demonstrate that using dual axis OPT, where two images are acquired simultaneously at different projection angles, can enable localization and tracking of features in 3-D with a time resolution equal to the camera frame rate. This is achieved with a 200x reduction in light dose compared to an equivalent volumetric time-lapse single camera OPT acquisition with 200 projection angles. We demonstrate the application of this technique to mapping the 3-D neutrophil migration pattern observed over ~25.5 minutes in a live 2 day post-fertilisation transgenic LysC:GFP zebrafish embryo following a tail wound.
authors.affiliation: Photonics Group, Department of Physics, Imperial College London, SW7 2AZ, UK ; These authors contributed equally to this work.
authors.forename: Lingling
authors.initials: L
authors.lastname: Chen
authors.affiliation: Photonics Group, Department of Physics, Imperial College London, SW7 2AZ, UK ; These authors contributed equally to this work.
authors.forename: Yuriy
authors.initials: Y
authors.lastname: Alexandrov
authors.affiliation: Photonics Group, Department of Physics, Imperial College London, SW7 2AZ, UK.
authors.forename: Sunil
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Institute for Chemical Biology, Department of Chemistry, imperial College London, SW7 2AZ, UK.
authors.forename: Natalie
authors.initials: N
authors.lastname: Andrews
authors.affiliation: Division of Cell and Molecular Biology, Department of Life Sciences, Imperial College London, SW7 2AZ, UK ; Centre for Integrative Systems Biology, Department of Life Sciences, Imperial College London, SW7 2AZ, UK.
authors.forename: Margaret J
authors.initials: MJ
authors.lastname: Dallman
authors.affiliation: Photonics Group, Department of Physics, Imperial College London, SW7 2AZ, UK ; These joint senior authors contributed equally to this paper.
authors.forename: Paul M W
authors.initials: PM
authors.lastname: French
authors.affiliation: Photonics Group, Department of Physics, Imperial College London, SW7 2AZ, UK ; These joint senior authors contributed equally to this paper.
authors.forename: James
authors.initials: J
authors.lastname: McGinty
keywords: (170.2520) Fluorescence microscopy; (170.3010) Image reconstruction techniques; (170.6900) Three-dimensional microscopy; (170.6920) Time-resolved imaging