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

Sci Rep. 2015 Nov 13:5:16625. doi: 10.1038/srep16625.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Disease Models, Animal
  • Dystrophin / deficiency
  • Dystrophin / genetics
  • Imaging, Three-Dimensional / methods
  • Larva / genetics
  • Larva / metabolism
  • Microscopy, Confocal
  • Microscopy, Phase-Contrast
  • Muscles / diagnostic imaging*
  • Muscular Dystrophy, Animal / diagnostic imaging
  • Muscular Dystrophy, Animal / genetics
  • Muscular Dystrophy, Duchenne / diagnostic imaging
  • Muscular Dystrophy, Duchenne / genetics
  • Myofibrils / diagnostic imaging*
  • Radiographic Image Enhancement / instrumentation
  • Radiographic Image Enhancement / methods*
  • Reproducibility of Results
  • Tomography, X-Ray Computed / instrumentation
  • Tomography, X-Ray Computed / methods*
  • Zebrafish

Substances

  • Dystrophin