Sepiolidae, Leach, 1817

Wild, Elvira, Wollesen, Tim, Haszprunar, Gerhard & Heß, Martin, 2015, Comparative 3 D microanatomy and histology of the eyes and central nervous systems in coleoid cephalopod hatchlings, Organisms Diversity & Evolution (New York, N. Y.) 15 (1), pp. 37-64 : 40

Family: Sepiolidae

Subfamily: Rossiinae R. macrosoma Delle Chiaje, 1829

Subfamily: Sepiolinae S. obscura Naef, 1916

Order: uncertain Family: Idiosepiidae I. notoides Berry, 1921

Order: Teuthoidea Suborder: Myopsida Family: Loliginidae L. vulgaris Lamarck, 1798

Superorder: Octopodiformes Order: Octopoda Suborder: Incirrata Family: Octopodidae O. vulgaris Cuvier, 1797

The hatchlings ( S. officinalis , L. vulgaris , I. notoides ) were fixed in 4 % formaldehyde in 0.1-M phosphate buffer (pH 7.4) overnight, and then, the heads were cut off and embedded in epoxy resin following standard protocols. Semithin section series (2.1–3 μm each, transversal planes, Table 3) were cut on an RMC MT-7000 microtome following a procedure established by Ruthensteiner (2008). Following a common convention, we use the physiological coordinate system for orientation instead of the morphological one, i.e., the mouth opening points forward (instead of ventrally), the funnel lies ventrally (not caudally). Hence, transversal sections are normal to the physiological anterior-posterior axis.

The sections were transferred to microscope slides, stained with Azur II-methylene blue (after Richardson et al. 1960), and mounted with coverslips. Single sections were photographed with an Olympus CX41 light microscope (PlanC N 10x or UPlan S Apo 40x objective) equipped with an Olympus DP25 digital camera, or complete microscope slides were photographed with an Olympus BX51-dotSlide system (UPlan S Apo 10x objective, XC 10 camera) using extended focal imaging and automated stitching.

The image series were converted from RGB color to grayscale, contrast enhanced, and unsharp masked with Adobe Photoshop 6.0 (San Jose, USA). Constrained by memory limitations of the imaging computer, the image stacks were partially reduced in size (see Table 3). The preprocessed images were loaded into Amira® 5.3.3 (Berlin, Germany), aligned, and segmented, i.e., the profiles of organs or other structures were selected manually in every image plane and attributed to separate “materials.” Therefrom, surfaces were rendered, smoothed, and displayed under any defined angle, color, transparency, and material composition. Appropriate views of the digital 3D models were virtually “photographed” using the snapshot tool and arranged to figures with Adobe Photoshop®. Interactive 3D models were inserted in the PDF figures following Ruthensteiner and Hess (2008) with updated software.