A 3D imaging and visualization workflow, using confocal microscopy and advanced image processing for brachyuran crab larvae

Kamanli, Seyit A.; Kihara, Terue C.; Ball, Alexander D.; Morritt, David; Clark, Paul F.

doi:10.1111/jmi.12540

Cited by 26 publications

(17 citation statements)

References 32 publications

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“…The study of internal morphology received a significant advance with the arrival of confocal laser scanning microscopy (CLSM), which uses stacks of two-dimensional sections to reconstruct three-dimensional models 12 , 13 . This approach has been applied in non-insect pancrustaceans for the study of embryos and larvae, but it is limited by light penetration and the staining properties of the tissues 14 – 16 .…”

Section: Introductionmentioning

confidence: 99%

Micro-computed tomography and histology to explore internal morphology in decapod larvae

Castejón

Alba‐Tercedor

Rotllant

et al. 2018

Sci Rep

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Traditionally, the internal morphology of crustacean larvae has been studied using destructive techniques such as dissection and microscopy. The present study combines advances in micro-computed tomography (micro-CT) and histology to study the internal morphology of decapod larvae, using the common spider crab (Maja brachydactyla Balss, 1922) as a model and resolving the individual limitations of these techniques. The synergy of micro-CT and histology allows the organs to be easily identified, revealing simultaneously the gross morphology (shape, size, and location) and histological organization (tissue arrangement and cell identification). Micro-CT shows mainly the exoskeleton, musculature, digestive and nervous systems, and secondarily the circulatory and respiratory systems, while histology distinguishes several cell types and confirms the organ identity. Micro-CT resolves a discrepancy in the literature regarding the nervous system of crab larvae. The major changes occur in the metamorphosis to the megalopa stage, specifically the formation of the gastric mill, the shortening of the abdominal nerve cord, the curving of the abdomen beneath the cephalothorax, and the development of functional pereiopods, pleopods, and lamellate gills. The combination of micro-CT and histology provides better results than either one alone.

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Section: Introductionmentioning

confidence: 99%

Micro-computed tomography and histology to explore internal morphology in decapod larvae

Castejón

Alba‐Tercedor

Rotllant

et al. 2018

Sci Rep

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“…Some image systems (e.g., SEM ) inevitably destroy type specimens; CLSM is highly desirable in this aspect as the studied specimen remains intact. According to Kamanli et al (2017) , the images obtained by CLSM are comparable in quality to SEM at the same magnifications, and the technique offers a 3D data set. In addition, the sample preparation routine for CLSM is simpler than that for SEM , it is practically a non-destructive method, and allows the study of hydrated material.…”

Section: Discussionmentioning

confidence: 99%

“…This is the first formal description of a marine copepod species based on combined use of interference and confocal microscopy in study of dissected appendages and the genital field. The methods for the acquisition of 3D rendered images are described by Corgosinho et al (2017) and Kamanli et al (2017) . One of the most important advantages of using Confocal Laser Scanning Microscopy ( CLSM ) over Scanning Electron Microscopy ( SEM ) is that CLSM is a nondestructive imaging technique for the study of whole microscopic animals or small parts of them, such as millimetre or micrometre-long hydrated structures (see extensive discussion by Kamanli et al 2017 ).…”

Section: Introductionmentioning

confidence: 99%

“…The methods for the acquisition of 3D rendered images are described by Corgosinho et al (2017) and Kamanli et al (2017) . One of the most important advantages of using Confocal Laser Scanning Microscopy ( CLSM ) over Scanning Electron Microscopy ( SEM ) is that CLSM is a nondestructive imaging technique for the study of whole microscopic animals or small parts of them, such as millimetre or micrometre-long hydrated structures (see extensive discussion by Kamanli et al 2017 ). In addition, the use of CLSM improves the presentation of appendages and structures in their natural 3-dimentional state, a property not easily transferable by the 2-dimentional inked drawings.…”

Section: Introductionmentioning

confidence: 99%

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Traditional and confocal descriptions of a new genus and two new species of deep water Cerviniinae Sars, 1903 from the Southern Atlantic and the Norwegian Sea: with a discussion on the use of digital media in taxonomy (Copepoda, Harpacticoida, Aegisthidae)

Corgosinho¹,

Kihara²,

Schizas³

et al. 2018

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Aegisthidae is one of the most abundant and diverse families of harpacticoid copepods living in deep-sea benthos, and the phylogenetic relationships within the family are in state of flux. Females of two new deep-water species of harpacticoid copepods belonging to the Hase gen. n. (Aegisthidae: Cerviniinae) are described. The first taxonomic description of marine copepod species based on the combined use of interference and confocal microscopy for the study of the habitus and dissected appendages is presented here. CLSM (Confocal Laser Scanning Microscopy) is a non-destructive method, comparable in quality to SEM (scanning electron microscopy) at the same magnifications. To observe and reconstruct in detail the habitus and dissected appendages, whole specimens and dissected parts were stained with Congo Red, mounted on slides with glycerine for CLSM and scanned under three visible-light lasers. Hase lagomorphicus gen. et sp. n. and Hase talpamorphicus gen. et sp. n. were collected from the sediments of the Southern Atlantic and the Norwegian Sea, from 2270 m and 5468 m depths, respectively. Hase gen. n. is included within Cerviniinae based on the caudal rami which are relatively divergent. Hase gen. n. is the sister taxon of Cerviniella based on the following synapomorphies: sturdy body, exopodites 1–3 of pereopods 1–3 heavily built, transformed into digging limbs, with strong outer and distal spines/setae, two-segmented endopod on the pereopods 2 and 3, and a reduced pereopod 5. Compared to Cerviniella, Hase gen. n. exhibits a more developed armature on the pereopod 1, which has outer and distal elements transformed into strong and long spines vs. stiff setae on Cerviniella.Hase gen. n. has one or two strong and long spines on the inner margin of the exopodite 3 of pereopod 4 and pereopod 5 is fused to the somite, ornamented with three distal setae. The telson of Hase gen. n. is subquadratic, and the furca is among the shortest yet described for Aegisthidae. The new species differ in a number of diagnostic characters, three of which are: a) the somite bearing pereopods 3 and 4 with latero-distal spiniform processes in H. talpamorphicus gen. et sp. n. but smooth in H. lagomorphicus gen. et sp. n., b) antenna is armed with three stout spines on the lateral inner margin of the exopod in H. talpamorphicus gen. et sp. n. and two proximal setae in H. lagomorphicus gen. et sp. n., and c) pereopod 4 exopodite 3 has two long and strong spines on the inner margin in H. lagomorphicus gen. et sp. n. and one spine in H. talpamorphicus gen. et sp. n. The high quality of CLSM images should foster discussion about the use of high quality digital images as type or as part of the type series in zoological studies, especially when studying rare and small macrofaunal and meiofaunal taxa.

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“…VGStudio Max (version 2.2) was initially used to view the samples immediately after the scanning by converting the data into TIFF stack formats in order to process the data using the open source 3D surface rendering program Drishti (Limaye 2012). The stack data stored as TIFF images were then imported using Drishti import (Ball et al 2017;Kamanli et al 2017;Koch et al 2017) and processed using Drishti. The most suitable positions of the images were taken as snapshots at different angles and saved in JPEG format.…”

Section: Methodsmentioning

confidence: 99%

Lobithelphusa mexicana Rodriguez, 1982 (Decapoda: Pseudothelphusidae): a reassessment of key characters and systematics

Villalobos-Hiriart

Kamanli

Álvarez

et al. 2019

Zootaxa

825

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Lobithelphusa Rodriguez, 1982 is endemic to southern Mexico. Assigned to this monotypic genus is the freshwater pseudothelphusid crab, L. mexicana Rodriguez, 1982. The original description based on an old, dry, male specimen in the collections of the Natural History Museum, London, is brief and the crab was only partially figured. Lobithelphusa mexicana has subsequently never been recorded from Mexico and, consequently, the conservation status of this species is unknown. The type specimen has a complete, but detached left chela; is devoid of the left ambulatory pereiopods 1–3, with the fourth missing the carpus, propodus and dactylus; and its left first gonopod detached but retained in a glass tube kept adjacent to the crab. In spite of this damage the crab is still in remarkably good condition for a specimen registered in 1860. The purpose of this study is to apply computed tomography (CT) scanning techniques on the extant type, including its first gonopod, in order to provide detailed illustrations of the crab. These, in turn, shall be used to re-describe the species in order to help establish the current distribution status of L. mexicana in Mexico, and clarify its systematic position within the Pseudothelphusidae Ortmann, 1893.

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A 3D imaging and visualization workflow, using confocal microscopy and advanced image processing for brachyuran crab larvae

Cited by 26 publications

References 32 publications

Micro-computed tomography and histology to explore internal morphology in decapod larvae

Micro-computed tomography and histology to explore internal morphology in decapod larvae

Traditional and confocal descriptions of a new genus and two new species of deep water Cerviniinae Sars, 1903 from the Southern Atlantic and the Norwegian Sea: with a discussion on the use of digital media in taxonomy (Copepoda, Harpacticoida, Aegisthidae)

Lobithelphusa mexicana Rodriguez, 1982 (Decapoda: Pseudothelphusidae): a reassessment of key characters and systematics

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