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.

中文翻译：

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改进的iDISCO方法对完整成人骨骼肌中肌内神经支配的三维可视化。

骨骼肌神经支配的三维可视化有助于理解形态结构和功能。iDISCO是一种完整的免疫标记和清除技术，为肌肉内神经纤维的体积成像提供了一种有价值的工具，但在使用鼠类一抗时会遭受由抗小鼠二抗引起的非特异性染色。通过引入称为m-iDISCO的ScaleCUBIC-1试剂的预处理，我们开发了一种改进的iDISCO方法。m-iDISCO方法可以消除非特异性染色，并使用小鼠抗神经丝初级抗体对各个肌肉中的神经纤维进行均匀，完整的标记。将m-iDISCO方法与光片显微镜相结合，使我们能够可视化成年小鼠胫前肌的神经支配，并从肌外分支到肌内终末分支追踪神经纤维。该方法是研究健康和疾病中完整骨骼肌神经支配的有效替代方法。