The histological processing involved sample fixation in dilute (1:2) Bouin’s fluid, dehydration in a graded ethanol series, clearing in xylene, paraffin-embedding (Histoplast, Argentina), and sectioning (5–10 μm-thick) on a rotary microtome (Microm). After staining with Gill’s hematoxylin-eosin, we examined and photographed the samples under a Nikon Eclipse 80i microscope using Nikon DS-Fi1-U3 camera and Nikon NIS-ELEMENT Image Software.
Also, we examined serially sectioned (10 μm-thick) Harris hematoxylin-eosin-stained lung samples under a Nikon Alphaphot-2 YS2 microscope equipped with a Nikon Digital Sight DS-5M camera. We selected the images of every fifth section for the 3D-reconstruction of the blood system and innervation of the lung. For this purpose, we used the software Reconstruct 1.1.0.0 (Fiala, 2005 (link)) and followed the procedure described by Ruthensteiner & Heß (2008) (link) to assemble the final PDF-model, as described in Rodriguez et al. (2019) (link).
Also, we examined serially sectioned (10 μm-thick) Harris hematoxylin-eosin-stained lung samples under a Nikon Alphaphot-2 YS2 microscope equipped with a Nikon Digital Sight DS-5M camera. We selected the images of every fifth section for the 3D-reconstruction of the blood system and innervation of the lung. For this purpose, we used the software Reconstruct 1.1.0.0 (Fiala, 2005 (link)) and followed the procedure described by Ruthensteiner & Heß (2008) (link) to assemble the final PDF-model, as described in Rodriguez et al. (2019) (link).
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