title: X-ray phase-contrast tomography for high-spatial-resolution zebrafish muscle imaging.
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.
authors.affiliation: Department of Applied Physics, KTH Royal Institute of Technology/Albanova, Stockholm, Sweden.
authors.forename: William
authors.initials: W
authors.lastname: Vågberg
authors.affiliation: Department of Applied Physics, KTH Royal Institute of Technology/Albanova, Stockholm, Sweden.
authors.forename: Daniel H
authors.initials: DH
authors.lastname: Larsson
authors.affiliation: Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Mei
authors.initials: M
authors.lastname: Li
authors.affiliation: Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Anders
authors.initials: A
authors.lastname: Arner
authors.affiliation: Department of Applied Physics, KTH Royal Institute of Technology/Albanova, Stockholm, Sweden.
authors.forename: Hans M
authors.initials: HM
authors.lastname: Hertz
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D016189:Dystrophin / Q000172:deficiency / Q000235:genetics; D021621:Imaging, Three-Dimensional / Q000379:methods; D007814:Larva / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D008858:Microscopy, Phase-Contrast; D009132:Muscles / Q000000981:diagnostic imaging*; D009137:Muscular Dystrophy, Animal / Q000000981:diagnostic imaging / Q000235:genetics; D020388:Muscular Dystrophy, Duchenne / Q000000981:diagnostic imaging / Q000235:genetics; D009210:Myofibrils / Q000000981:diagnostic imaging*; D011856:Radiographic Image Enhancement / Q000295:instrumentation / Q000379:methods*; D015203:Reproducibility of Results; D014057:Tomography, X-Ray Computed / Q000295:instrumentation / Q000379:methods*; D015027:Zebrafish
chemical_list: D016189:Dystrophin