Mesoscopic in vivo 3-D tracking of sparse cell populations using angular multiplexed optical projection tomography
- 12 March 2015
- journal article
- Published by Optica Publishing Group in Biomedical Optics Express
- Vol. 6 (4), 1253-61
- https://doi.org/10.1364/boe.6.001253
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.Keywords
Funding Information
- Biotechnology and Biological Sciences Research Council (BBSRC) (BB/L018039/1)
- European Foundation for the Study of Diabetes (EFSD)
- Medical Research Council (MRC) (MR/K011561/1)
- Wellcome Trust (WT 095931/Z/11/Z)
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