1. Three-dimensional visualization of intramuscular innervation in intact adult skeletal muscle by a modified iDISCO method
- Author
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Dan Zhu, Jingtan Zhu, Jianyi Xu, Yusha Li, and Tingting Yu
- Subjects
Paper ,Neuroscience (miscellaneous) ,01 natural sciences ,010309 optics ,whole-mount immunostaining ,03 medical and health sciences ,Immunolabeling ,0302 clinical medicine ,0103 physical sciences ,medicine ,Radiology, Nuclear Medicine and imaging ,skeletal muscle ,Radiological and Ultrasound Technology ,biology ,Chemistry ,3d image processing ,Skeletal muscle ,Anatomy ,Primary and secondary antibodies ,Research Papers ,Structure and function ,Staining ,medicine.anatomical_structure ,Three dimensional visualization ,biology.protein ,030217 neurology & neurosurgery ,intramuscular innervation - Abstract
Three-dimensional visualization of the innervation in skeletal muscles is helpful for understanding the morphological structure and function. iDISCO, a whole-mount immunolabeling and clearing technique, provides a valuable tool for volume imaging of intramuscular nerve fibers but suffers from the nonspecific staining caused by the anti-mouse secondary antibody when using the murine primary antibody. We developed a modified iDISCO method by introducing pretreatment of ScaleCUBIC-1 reagent, termed m-iDISCO. The m-iDISCO method could eliminate the nonspecific staining and achieve uniform and complete labeling of nerve fibers in various muscles with mouse anti-neurofilament primary antibody. Combining the m-iDISCO method with light-sheet microscopy enabled us to visualize the innervation of adult mouse tibialis anterior and trace the nerve fibers from extramuscular branches to intramuscular terminal branches. This method represents an effective alternative for studying the innervation of intact skeletal muscles in health and disease.
- Published
- 2019