Many techniques are used today to study insect morphology, including light and electron microscopy. Most of them require to specifically prepare the sample, precluding its use for further investigation. In contrast, micro-CT allows a sample to be studied in a non-destructive and rapid process, even without specific treatments that might hinder the use of rare and hard-to-find species in nature. We used synchrotron radiation (SR) micro-CT and conventional micro-CT to prepare 3D reconstructions of Diptera, Coleoptera, and Hymenoptera species that had been processed with 4 common preparation procedures: critical-point drying, sputter-coating, resin embedding, and air-drying. Our results showed that it is possible to further utilize insect samples prepared with the aforementioned preparation techniques for the creation of 3D models. Specimens dried at the critical point showed the best results, allowing us to faithfully reconstruct both their external surface and their internal structures, while sputter-coated insects were the most troublesome for the 3D reconstruction procedure. Air-dried specimens were suitable for external morphological analyses, while anatomical investigation of soft internal organs was not possible due to their shrinking and collapsing. The sample included in resin allowed us to reconstruct and appreciate the external cuticle and the internal parts. In this work, we demonstrate that insect samples destined to different analyses can be used for new micro-CT studies, further deepening the possibility of state-of-the-art morphological analyses.

The genus Paussus is a highly specialized, charismatic group of ground beetles (Carabidae) classified in the subfamily Paussinae. All species of Paussus are obligate myrmecophiles (associates of ants). As with many other myrmecophilous or termitophilous beetles, Paussus have undergone extreme phenotypic adaptations for life with ants, at the level of head, antennae, and prothorax. Host data suggest that Paussus species are likely to be species-specific ant parasites, and the structural modifications of antennae and other body parts are likely under selection by their host ants. Investigating anatomical structures have been fundamental to better understand living organisms, and their interplay with the surrounding environment, which could induce significant morphological variation. In the last few years, bio-imaging techniques paired with geometric morphometrics (GM) overcame the limits of traditional anatomical studies, becoming widely non-invasive and highly informative for both internal and external characters. The use of Computed Tomography (CT) scanners definitively allowed to advance in the knowledge of either known or neglected biological structures. For this project, we used X-ray micro-computed ‡ ‡ ‡