X-ray phase-contrast tomography for high-spatial-resolution zebrafish muscle imaging

• Published in: Scientific reports 2015-11, Vol.5 (1), p.16625-16625, Article 16625
• Main Authors: Vågberg, William, Larsson, Daniel H., Li, Mei, Arner, Anders, Hertz, Hans M.
• Format: Article
• Language: English
• Subjects: 14/63; 59; 631/1647/245/1847; 639/166/985; Animal models; Animals; Disease Models, Animal; Duchenne's muscular dystrophy; Dystrophin; Dystrophin - deficiency; Dystrophin - genetics; Dystrophy; Fertilization; Histology; Humanities and Social Sciences; Imaging, Three-Dimensional - methods; Larva - genetics; Larva - metabolism; Larvae; Medical imaging; Microscopy, Confocal; Microscopy, Phase-Contrast; multidisciplinary; Muscles - diagnostic imaging; Muscular Dystrophy, Animal - diagnostic imaging; Muscular Dystrophy, Animal - genetics; Muscular Dystrophy, Duchenne - diagnostic imaging; Muscular Dystrophy, Duchenne - genetics; Myofibrils; Myofibrils - diagnostic imaging; Radiographic Image Enhancement - instrumentation; Radiographic Image Enhancement - methods; Reproducibility of Results; Science; Spatial discrimination; Tomography; Tomography, X-Ray Computed - instrumentation; Tomography, X-Ray Computed - methods; Zebrafish
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep16625

Imaging of muscular structure with cellular or subcellular detail in whole-body animal models is of key importance for understanding muscular disease and assessing interventions. Classical histological methods for high-resolution imaging methods require excision, fixation and staining. Here we show that the three-dimensional muscular structure of unstained whole zebrafish can be imaged with sub-5 μm detail with X-ray phase-contrast tomography. Our method relies on a laboratory propagation-based phase-contrast system tailored for detection of low-contrast 4–6 μm subcellular myofibrils. The method is demonstrated on 20 days post fertilization zebrafish larvae and comparative histology confirms that we resolve individual myofibrils in the whole-body animal. X-ray imaging of healthy zebrafish show the expected structured muscle pattern while specimen with a dystrophin deficiency ( sapje ) displays an unstructured pattern, typical of Duchenne muscular dystrophy. The method opens up for whole-body imaging with sub-cellular detail also of other types of soft tissue and in different animal models.