Whole-organism 3D quantitative characterization of zebrafish melanin by silver deposition micro-CT.
Whole-organism 3D quantitative characterization of zebrafish melanin by silver deposition micro-CT.
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DOI：
10.7554/elife.68920
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发表时间：
2021-09-16
期刊：
影响因子：
7.7
通讯作者：
Cheng KC
中科院分区：
文献类型：
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作者：
Katz SR;Yakovlev MA;Vanselow DJ;Ding Y;Lin AY;Parkinson DY;Wang Y;Canfield VA;Ang KC;Cheng KC
我们先前描述了X射线组织断层扫描，一种高分辨率、非破坏性形式的X射线显微断层扫描（micro-CT）成像，其定制用于三维（3D）、数字组织学，允许定量、体积组织和生物体表型分析（Ding等人，2019年）。在这里，我们结合了微CT与离子银染色的一种新的应用，以表征整个斑马鱼幼虫的黑色素分布。由此产生的图像能够对局部黑色素含量和形态进行全身计算分析。银染鱼的标准化micro-CT重建一致地再现了光学显微镜观察到的色素模式，并进一步允许对野生型和突变体样品中的黑色素含量进行直接定量比较，包括以前未注意到的细微表型。用于微CT的黑色素银染色提供了以细胞分辨率对特定细胞类型进行全身、3D计算表型组学分析的原理证明，具有在其他模式生物和黑色素细胞肿瘤中的潜在应用。在全生物体、高分辨率表型分析方面的进展为研究遗传、疾病和环境变量的表型效应提供了上级背景。
We previously described X-ray histotomography, a high-resolution, non-destructive form of X-ray microtomography (micro-CT) imaging customized for three-dimensional (3D), digital histology, allowing quantitative, volumetric tissue and organismal phenotyping (Ding et al., 2019). Here, we have combined micro-CT with a novel application of ionic silver staining to characterize melanin distribution in whole zebrafish larvae. The resulting images enabled whole-body, computational analyses of regional melanin content and morphology. Normalized micro-CT reconstructions of silver-stained fish consistently reproduced pigment patterns seen by light microscopy, and further allowed direct quantitative comparisons of melanin content across wild-type and mutant samples, including subtle phenotypes not previously noticed. Silver staining of melanin for micro-CT provides proof-of-principle for whole-body, 3D computational phenomic analysis of a specific cell type at cellular resolution, with potential applications in other model organisms and melanocytic neoplasms. Advances such as this in whole-organism, high-resolution phenotyping provide superior context for studying the phenotypic effects of genetic, disease, and environmental variables.