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diceCT: a valuable technique to study the nervous system of fishes
journal contributionposted on 2021-01-06, 01:30 authored by Victoria Camilieri-Asch, Jeremy A Shaw, Andrew Mehnert, Kara E Yopak, Julian C Partridge, Shaun CollinShaun Collin
© 2020 Camilieri-Asch et al. Contrast-enhanced X-ray imaging provides a non-destructive and flexible approach to optimizing contrast in soft tissues, especially when incorporated with Lugol’s solution (aqueous I2KI), a technique currently referred to as diffusible iodine-based contrast-enhanced computed tomography (diceCT). This stain exhibits high rates of penetration and results in excellent contrast between and within soft tissues, including the central nervous system. Here, we present a staining method for optimizing contrast in the brain of a cartilaginous fish, the brownbanded bamboo shark, Chiloscyllium punctatum, and a bony fish, the common goldfish, Carassius auratus, usingdiceCT.Theaim of this optimization procedure is to provide suitable contrast between neural tissue and background tissue(s) of the head, thereby facilitating digital segmentation and volumetric analysis of the central nervous system. Both species were scanned before staining and were rescanned at time (T) intervals, either every 48 h (C. punctatum) or every 24 h (C. auratus), to assess stain penetration and contrast enhancement. To compare stain intensities, raw X-ray CT data were reconstructed using air and water calibration phantoms that were scanned under identical conditions to the samples. Optimal contrast across the brain was achieved at T = 240 h for C. punctatum and T=96 h for C. auratus. Higher resolution scans of the whole brain were obtained at the two optimized staining times for all the corresponding specimens. The use of diceCT provides a new and valuable tool for visualizing differences in the anatomic organization of both the central and peripheral nervous systems of fish.
V.C.A. was supported by the Australian Federal Government and The University of Western Australia (UWA) in the form of an Australian Training Program and The UWA Safety-Net Top-up scholarships.
Article NumberARTN 0076-20.2020
PublisherSociety for Neuroscience
Rights StatementThe Author reserves all moral rights over the deposited text and must be credited if any re-use occurs. Documents deposited in OPAL are the Open Access versions of outputs published elsewhere. Changes resulting from the publishing process may therefore not be reflected in this document. The final published version may be obtained via the publisher’s DOI. Please note that additional copyright and access restrictions may apply to the published version.
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Science & TechnologyLife Sciences & BiomedicineNeurosciencesNeurosciences & NeurologydiceCTcomparative neuroanatomyelasmobranchteleostiodineX-ray tomographyCOMPUTED-TOMOGRAPHY DICECTMICRO-CTBRAIN ORGANIZATIONQUANTITATIVE-ANALYSISVOLUMETRIC-ANALYSISONTOGENIC SHIFTSCONTRAST AGENTSHIGH-RESOLUTIONTISSUEMORPHOLOGY