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. 2017 Jun 7;7:2917. doi: 10.1038/s41598-017-03050-y

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Respiratory and anaesthesia perfusion set-up for adult zebrafish in vivo imaging. (a) Schematic drawing of the perfusion system and MRI set-up. IFT – inflow tube, OFS – outflow sink, OFT – outflow tube, PS - pressure sensor (b) Ventral view (anterior at the top) of a zebrafish showing an intra-peritoneal injection of contrast agent prior to MRI. (c) Photograph showing the thin transparent respiratory perfusion inflow tube (IFT, arrow) positioned in the zebrafish mouth. Note the water level is above the eyes of the fish and covers the gills. (d) Drawing of the fish perfusion chamber as used for 3D printing illustrating the contours of the fish well. (e) Empty fish well of the perfusion chamber (view from top) with the outflow sink visible. (f) Fish well containing an adult zebrafish with perfusion inflow tube (IFT, arrow) in its mouth ready for imaging (see also c). Note the fish is 2/3 covered with water and eyes and gills are submerged. (g) Whole perfusion chamber (top view) containing a fish with and without the lid. The perfusion chamber is screwed onto a larger tube (dark grey) to insert it into the MR magnet. (h–i) Single image plane of a 3D image stack of a live adult zebrafish heart (h) or the whole scan field (i) from eye to mid swim bladder in (sagittal view). Scale bar 1mm. (j) Single image plane of a 3D image stack of a paraformaldehyde-fixed adult zebrafish in sagittal view. a-atrium, b-bulbus arteriosus, e-eye, i-intestine, k-kidney, l-liver, m-muscle, n-olfactory pit, o-oesophagus, p-pharynx, s-swim bladder, t-tectum opticum, v-ventricle.