Whole-organism 3D quantitative characterization of zebrafish melanin by silver deposition micro-CT

eLife

Dilworth Y Parkinson¹  Victor A Canfield²  Alex Y Lin²  Maksim A Yakovlev²  Daniel J Vanselow²  Yifu Ding³  Spencer R Katz³  Khai C Ang⁴  Keith C Cheng⁴  Yuxin Wang⁵ 

[1] Advanced Light Source, Lawrence Berkeley National Labs, Berkeley, United States;Division of Experimental Pathology, Department of Pathology, Pennsylvania State University College of Medicine, Hershey, United States;The Jake Gittlen Laboratories for Cancer Research, Penn State College of Medicine, Hershey, United States;Division of Experimental Pathology, Department of Pathology, Pennsylvania State University College of Medicine, Hershey, United States;The Jake Gittlen Laboratories for Cancer Research, Penn State College of Medicine, Hershey, United States;Medical Scientist Training Program, Penn State College of Medicine, Hershey, United States;Division of Experimental Pathology, Department of Pathology, Pennsylvania State University College of Medicine, Hershey, United States;The Jake Gittlen Laboratories for Cancer Research, Penn State College of Medicine, Hershey, United States;Zebrafish Functional Genomics Core, Penn State College of Medicine, Hershey, United States;Mobile Imaging Innovations, Inc, Palatine, United States;

关键词: melanin;    pigmentation;    micro-CT;    silver staining;    3D imaging;    whole-organism phenomics;    Zebrafish;   

DOI  :  10.7554/eLife.68920

来源: eLife Sciences Publications, Ltd

【 摘 要 】

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.

【 授权许可】

CC BY