Fig. 1.

Whole-tissue immunolabeling of the intact mouse femurs and hindpaws to image neural anatomy and pathology. a, b The intact, unsectioned femurs of adult mice before (a) and after (b) BoneClear. c–e The femurs were processed for the whole-tissue immunolabeling of PGP9.5 (c), TH (d) or GFAP (e). Representative 3D-projection images at 1.26× magnification on the lightsheet microscope (left panels), and representative 3D-projection images of 300 μm-depth bone marrow in the proximal (upper-right panels) and middle (lower-right panels) shafts at 20× magnification on the confocal microscope were shown. f–h Ablation of the intraosseous sympathetic innervations in the femurs of Th-Cre; TrkA^fl/fl mice. The femurs were processed for the immunolabeling of TH (f) or PGP9.5 (g). Representative 3D-projection images at 1.26× magnification on the lightsheet microscope were shown. The length of the TH-positive sympathetic axons in the femurs was quantified (h). n = 4, mean ± SEM, *P < 0.01 (Student’s t-test). i, j Chemotherapy-induced neuropathy of the intraosseous sympathetic innervations. The femurs of the vehicle-treated (left panel) or Paclitaxel-treated (right panel) mice were processed for the TH immunolabeling. Representative 3D-projection images at 1.26× magnification on the lightsheet microscope were shown (i). The length of the TH-positive sympathetic axons in the femurs was quantified (j). n = 4, mean ± SEM, *P < 0.01 (Student’s t-test). k, l The intact, unsectioned hindpaws of adult mice before (k) and after (l) BoneClear. m The hindpaw was processed for the whole-tissue PGP9.5 immunolabeling. n, o Traumatic injury-induced neuropathy in the hindpaw. Sarm1^+/+ (n) or Sarm1^−/− (o) mice were subjected to the sciatic nerve injury. The hindpaws were processed for the PGP9.5 immunolabeling at 6 days post injury. p Post-injury neural regeneration in the hindpaw. The hindpaws of wild-type mice were processed for the PGP9.5 immunolabeling at 2 weeks (left panel) or 4 weeks (right panel) after the sciatic nerve injury. q, r Chemotherapy-induced neuropathy in the hindpaw. Sarm1^+/+ (q) or Sarm1^−/− (r) mice were treated with Paclitaxel for 6 days, and the hindpaws were processed for the PGP9.5 immunolabeling. Representative 3D-projection images at 1.26× magnification on the lightsheet microscope (left panels, the squares denote the skin regions imaged at 12.6× magnification), and representative 3D-projection images of the nail (upper-right panels), digit tip (middle-right panels) and 300 μm-depth skin (lower-right panels) at 12.6× magnification were shown (m, n, o, q, r)