Synchrotron X-ray phase contrast micro tomography to explore the morphology of abdominal organs in Pterostichus melas italicus Dejean, 1828 (Coleoptera, Carabidae)

Exploring Compound Eyes in Adults of Four Coleopteran Species Using Synchrotron X-ray Phase-Contrast Microtomography (SR-PhC Micro-CT)

“…Males and females from each species were anaesthetised in a cold chamber at 4 °C for three minutes and prepared as indicated in [ 6 ]. Briefly, beetles were fixed in 2.5% glutaraldehyde and 1% paraformaldehyde in 0.1 M phosphate buffer, at pH 7.4 (PBS; Electron Microscopy Sciences), overnight at 4 °C, washed with PBS and dehydrated in a graded ethanol series.…”

“…This method avoids artefacts resulting from invasive dissections, followed by relatively time-consuming fixing and physical tissue slicing, required for image analyses under light and electron microscopy. Indeed, micro-CT has proven to be useful for virtual dissections, 3D reconstruction and morphological descriptions of the head [ 1 , 2 ], muscles [ 2 , 3 , 4 ], brain [ 5 ], digestive [ 6 , 7 ] and reproductive [ 8 , 9 , 10 , 11 ] systems, as well as insect fossils [ 12 , 13 , 14 , 15 ]. Moreover, Synchrotron radiation X-ray phase-contrast microtomography (SR-PhC micro-CT) allows the use of high-resolution imaging coupled with segmentation, for 3D morphological analyses with high image contrast-to-noise ratios in biological tissues, and does not require the use of contrast agents, even in samples with weak X-ray absorption [ 16 , 17 , 18 , 19 ].…”

“…Moreover, Synchrotron radiation X-ray phase-contrast microtomography (SR-PhC micro-CT) allows the use of high-resolution imaging coupled with segmentation, for 3D morphological analyses with high image contrast-to-noise ratios in biological tissues, and does not require the use of contrast agents, even in samples with weak X-ray absorption [ 16 , 17 , 18 , 19 ]. In recent decades, it has been applied as a non-invasive technique to observe external and internal anatomical structures of living insects [ 20 , 21 ], and specimens immersed in ethanol after fixation [ 2 , 6 ] or embedded in amber [ 22 ].…”

Anatomical study of the red flour beetle using synchrotron radiation X‐ray phase‐contrast micro‐tomography

“…Synchrotron Radiation X‐ray phase‐contrast microtomography (SR‐PhC micro‐CT) is a powerful imaging technique for the study of anatomical structures and, when followed by an appropriate volumetric image segmentation, for 3D morphological analyses using an enhanced contrast‐to‐noise ratio in biological tissues, and does not require the use of contrast agents even in samples with weak X‐ray absorption (Donato et al, 2017; Elfarnawany et al, 2017; Endrizzi et al, 2018; Hellerhoff et al, 2019; Lak et al, 2008; Oliva et al, 2020). In previous entomological studies, virtual dissections performed using microtomography (micro‐CT) have provided morphological descriptions of various anatomical parts such as head (Betz et al, 2007; Hörnschemeyer et al, 2002), muscles (Bäumler et al, 2018; Li et al, 2011), brain (Smith et al, 2016), digestive (Alba‐Alejandre et al, 2019; Donato et al, 2021) visual (Giglio et al, 2022), and reproductive (Alba‐Alejandre et al, 2020; Küpper et al, 2019; Vommaro et al, 2022) systems. This non‐invasive method enhances the visualization of structures in situ and avoids all the artefacts associated with dissections and preparation of histological sections using traditional histological techniques, reducing the time of sample preparation protocols (Betz et al, 2007; Donato et al, 2021; Westneat et al, 2008).…”

“…In previous entomological studies, virtual dissections performed using microtomography (micro‐CT) have provided morphological descriptions of various anatomical parts such as head (Betz et al, 2007; Hörnschemeyer et al, 2002), muscles (Bäumler et al, 2018; Li et al, 2011), brain (Smith et al, 2016), digestive (Alba‐Alejandre et al, 2019; Donato et al, 2021) visual (Giglio et al, 2022), and reproductive (Alba‐Alejandre et al, 2020; Küpper et al, 2019; Vommaro et al, 2022) systems. This non‐invasive method enhances the visualization of structures in situ and avoids all the artefacts associated with dissections and preparation of histological sections using traditional histological techniques, reducing the time of sample preparation protocols (Betz et al, 2007; Donato et al, 2021; Westneat et al, 2008). Three‐dimensional (3D) rendering is useful for studying the volumetric arrangement of organs in miniature insects (Alba‐Alejandre et al, 2019), for toxicological applications (Smith et al, 2020) or for creating data sets of the internal anatomy of model insects (Rother et al, 2021).…”