To produce the precise three-dimensional geometry of the AT for the mathematical models and for morphological measurements, six AT of P. cieloi (three male and three female) were scanned using a SkyScan 1172 X-ray μ-CT scanner (Bruker Corporation, Billerica, MA, USA) with a resolution of 12.9 μm (50 kV source voltage, 200 μA source current, 200 ms exposure and 0.2° rotation steps). For obtaining a series of orthogonal slices, the μ-CT projection images were reconstructed with NRecon (v. 1.6.9.18, Bruker Corporation, Billerica, MA, USA).
The three-dimensional segmentation of the ATs were performed with the software Amira-Aviso 6.7 (Thermo Fisher Scientific, Waltham, MA, USA) and exported as stereolithography (STL) files for numerical modelling using established protocols [12 (link),14 (link)]. The segmented images were also used for obtaining the AT cross-sectional radius, length and volume measurements through the Center Line Tree module in AMIRA. For the two-dimensional measurement of spiracle size, an Alicona InfiniteFocus microscope (G5, Bruker Alicona Imaging, Graz, Austria) at ×5 objective magnification was used, with a resolution of about 100 nm.
The three-dimensional segmentation of the ATs were performed with the software Amira-Aviso 6.7 (Thermo Fisher Scientific, Waltham, MA, USA) and exported as stereolithography (STL) files for numerical modelling using established protocols [12 (link),14 (link)]. The segmented images were also used for obtaining the AT cross-sectional radius, length and volume measurements through the Center Line Tree module in AMIRA. For the two-dimensional measurement of spiracle size, an Alicona InfiniteFocus microscope (G5, Bruker Alicona Imaging, Graz, Austria) at ×5 objective magnification was used, with a resolution of about 100 nm.
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