The use of micro-computed tomography as a minimally invasive tool for anatomical study of bivalves (Mollusca: Bivalvia)

Machado, Fabrizio Marcondes; Passos, Flávio Dias; Giribet, Gonzalo

doi:10.1093/zoolinnean/zly054

Cited by 25 publications

(10 citation statements)

References 37 publications

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“…Many polychaete descriptions are traditionally based on stereomicroscopy, frequently supported by whole body optical microscopy and SEM, while micro-dissection and histology has normally been applied for purely anatomical studies (see above). However, the use of micro-CT for species identification in other taxa seems particularly relevant since it avoids specimen destruction, which is inevitable after histological sectioning (Fernández et al, 2014; Pedrouzo et al, 2017; Machado, Passos & Giribet, 2018). This is an evident advantage of micro-CT over HIS, although the former has obvious limitations to properly visualize many characters, especially at cellular level and, therefore, classical histological techniques are still essential for descriptions of new species when a proper identification of internal structures is required.…”

Section: Discussionmentioning

confidence: 99%

An integrative approach to the anatomy of Syllis gracilis Grube, 1840 (Annelida) using micro-computed X-ray tomography

Parapar

Caramelo

Candás

et al. 2019

PeerJ

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Background The overall anatomy of the genus Syllis (Annelida: Syllidae) has been largely studied; however, an integrative approach considering different anatomical techniques has never been considered. Here, we use micro-computed X-ray tomography (micro-CT) to examine the internal anatomy of Syllis gracilis Grube, 1840, along with other widely available techniques. Methods We studied the anatomy of the marine annelid S. gracilis through an integrative approach, including micro-CT along with stereo and light compound microscopy (STM, LCM), scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM) and histological sectioning (HIS). In this manner, we evaluated the applicability of micro-CT for the examination of annelid anatomy by testing whether the images obtained make it possible to visualize the main body structures, in comparison with other current techniques, of the various elements of its internal anatomy. Results Overall external and internal body elements are clearly shown by the integrative use of all techniques, thus overcoming the limitations of each when studied separately.Any given method shows disparate results, depending on the body part considered. For instance, micro-CT provided good images of the external anatomy, including relevant characters such as the shape, length and number of articles of dorsal parapodial cirri. However, it is especially useful for the examination of internal anatomy, thus allowing for 3D visualization of the natural spatial arrangement of the different organs. The features best visualized are those of higher tissue density (i.e., body musculature, anterior parts of the digestive tract), particularly in 3D images of unstained specimens, whereas less electrodense tissues (i.e., the peritoneal lining of septa and nervous system) are less clearly visualized. The use of iodine stain with micro-CT has shown advantages against non-staining for the adequate observation of delicate elements of low density, such as the segmental organs, the connective between the ganglia, the ventral nerve cord and segmental nerves. Discussion Main external anatomical elements of S. gracilis are well shown with micro-CT, but images show lesser optical resolution and contrast when compared to micrographs provided by SEM and CLSM, especially for fine structural features of chaetae. Comparison of micro-CT and HIS images revealed the utility and reliability of the former to show the presence, shape and spatial disposition of most internal body organs; the resolution of micro-CT images at a cellular level is, however, much lower than that of HIS, which makes both techniques complementary.

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Section: Discussionmentioning

confidence: 99%

An integrative approach to the anatomy of Syllis gracilis Grube, 1840 (Annelida) using micro-computed X-ray tomography

Parapar

Caramelo

Candás

et al. 2019

PeerJ

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“…The improving speed and resolution of µ-CT scanning enables increasingly focussed morphological studies targeting specific organs and tissues, and in smaller subjects 13–17 . Initial studies on molluscs have also proven to be useful for examination of anatomical details in bivalves 18 . Of particular interest to this study is the soft-tissue visualisation and imaging in invertebrates, including central nervous and sensory systems 2,12,19–23 .…”

Section: Introductionmentioning

confidence: 99%

The utility of micro-computed tomography for the non-destructive study of eye microstructure in snails

Sumner‐Rooney

Kenny

Ahmed

et al. 2019

Sci Rep

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Molluscan eyes exhibit an enormous range of morphological variation, ranging from tiny pigment-cup eyes in limpets, compound eyes in ark clams and pinhole eyes in Nautilus, through to concave mirror eyes in scallops and the large camera-type eyes of the more derived cephalopods. Here we assess the potential of non-destructive micro-computed tomography (µ-CT) for investigating the anatomy of molluscan eyes in three species of the family Solariellidae, a group of small, deep-sea gastropods. We compare our results directly with those from traditional histological methods applied to the same specimens, and show not only that eye microstructure can be visualised in sufficient detail for meaningful comparison even in very small animals, but also that μ-CT can provide additional insight into gross neuroanatomy without damaging rare and precious specimens. Data from μ-CT scans also show that neurological innervation of eyes is reduced in dark-adapted snails when compared with the innervation of cephalic tentacles, which are involved in mechanoreception and possibly chemoreception. Molecular tests also show that the use of µ-CT and phosphotungstic acid stain do not prevent successful downstream DNA extraction, PCR amplification or sequencing. The use of µ-CT methods is therefore highly recommended for the investigation of difficult-to-collect or unique specimens.

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“…This possibility has so far been used to replace existing, invasive techniques or was carried out to complement data derived from more conventional modalities. Using contrast-enhanced µCT, new species have been described in a number of metazoan taxa, including Myriapoda (Akkari et al 2015), Hirudinea (Tessler et al 2016), Gastropoda (Jochum et al 2017), Hymenoptera (Garcia et al 2017), Bivalvia (Machado et al 2018), and Cnidaria (Gusmão et al 2018). Although specific CT techniques have recently also been used in echinoderm taxonomy (Mihaljević & Rosenblatt 2018, Souto & Martins 2018, these studies focused on echinoderm hard part structures only.…”

Section: Morphological and Anatomical Aspectsmentioning

confidence: 99%

<p class="HeadingRunIn"><strong>Combined visualization of echinoderm hard and soft parts using contrast-enhanced micro-computed tomography</strong></p>

Ziegler

2019

Zoosymposia

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Recent studies have shown that micro-computed tomography (µCT) must be considered one of the most suitable techniques for the non-invasive, three-dimensional (3D) visualization of metazoan hard parts. In addition, µCT can also be used to visualize soft part anatomy non-destructively and in 3D. In order to achieve soft tissue contrast using µCT based on X-ray attenuation, fixed specimens must be immersed in staining solutions that include heavy metals such as silver (Ag), molybdenum (Mo), osmium (Os), lead (Pb), or tungsten (W). However, while contrast-enhancement has been successfully applied to specimens pertaining to various higher metazoan taxa, echinoderms have thus far not been analyzed using this approach. In order to demonstrate that this group of marine invertebrates is suitable for contrast-enhanced µCT as well, the present study provides results from an application of this technique to representative species from all five extant higher echinoderm taxa. To achieve soft part contrast, freshly fixed and museum specimens were immersed in an ethanol solution containing phosphotungstic acid and then scanned using a high-resolution desktop µCT system. The acquired datasets show that the combined visualization of echinoderm soft and hard parts can be readily accomplished using contrast-enhanced µCT in all extant echinoderm taxa. The results are compared with µCT data obtained using unstained specimens, with conventional histological sections, and with data previously acquired using magnetic resonance imaging, a technique known to provide excellent soft tissue contrast despite certain limitations. The suitability for 3D visualization and modeling of datasets gathered using contrast-enhanced µCT is illustrated and applications of this novel approach in echinoderm research are discussed.

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The use of micro-computed tomography as a minimally invasive tool for anatomical study of bivalves (Mollusca: Bivalvia)

Cited by 25 publications

References 37 publications

An integrative approach to the anatomy of Syllis gracilis Grube, 1840 (Annelida) using micro-computed X-ray tomography

An integrative approach to the anatomy of Syllis gracilis Grube, 1840 (Annelida) using micro-computed X-ray tomography

The utility of micro-computed tomography for the non-destructive study of eye microstructure in snails

<p class="HeadingRunIn"><strong>Combined visualization of echinoderm hard and soft parts using contrast-enhanced micro-computed tomography</strong></p>

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