Dikerogammarus reconstruction

Reier, Susanne, Sattmann, Helmut, Schwaha, Thomas, Harl, Josef, Konecny, Robert & Haring, Elisabeth, 2019, An integrative taxonomic approach to reveal the status of the genus Pomphorhynchus Monticelli, 1905 (Acanthocephala: Pomphorhynchidae) in Austria, International Journal for Parasitology: Parasites and Wildlife 8, pp. 145-155 : 146

2.2.2. Sectioning and 3- D reconstruction

To reconstruct important morphological characters such as the extensions of the basal hooks, 3-D reconstructions of histological semithin sections were performed. Therefore, probosces of the different species were cut from the trunk and rapidly dehydrated via acidified dimethoxypropane (DMP). Then they were embedded into Agar LVR resin (Agar Scientific, Stansted, Essex, UK) via acetone as intermediate. Serial semithin sections of cured resin blocks were prepared with a Leica UC6 ultramicrotome (Leica Microsystems, Wetzlar, Germany) at a thickness of 1 μm. Sections were stained with toluidine blue and subsequently examined on a Nikon NiU light microscope (Nikon Instruments, Tokyo, Japan) equipped with a Ds-Ri2 microscope camera. Image stacks from the serial sections were converted to greyscales and imported into the 3 D reconstruction software Amira 6.11 (FEI, Hillsboro, Oregon, USA). The general outline of the proboscis was displayed by using a volume rendering of the grey-scale information contained in the histological series ( Handschuh et al., 2010). Segmentation of single hooks was conducted manually with the brush tool. A surface of the segmented hooks was generated ( Ruthensteiner, 2008) and combined with the volume rendering for a general outline. Snapshots were taken with the Amira 6.11 software (FEI, Hillsboro, Oregon, USA). At least one longitudinal row of hooks was reconstructed for each of the five specimens.