Fig. 9
From: Tissue clearing of both hard and soft tissue organs with the PEGASOS method
Nerves and arteries have distinct distribution patterns within the marrow space of long bone. a Thoracic cage from adult Wnt1-Cre;Ai14 mouse (2-months-old) was cleared following the PEGASOS recirculation procedure and imaged with a two-photon microscope to display thoracic vertebrae, ribs, sympathetic trunks, and spinal nerves (arrows) exiting in the intervertebral foramina. b Dotted box in a is enlarged to display sympathetic ganglion (SG), DRG and the communication ramus (arrows). c Nerve bundles in the femur periosteum. d Maximum Z-projection of the tibia (~1.5 mm thickness) imaged with a 10× objective shows the innervation within the bone marrow. Dotted areas are enlarged in e–h. Arrows in g and h indicate nerve fibers penetrating the cortical bone. An intact tibia from αSMA-CreERT2;Ai14 mice was cleared and imaged with a two-photon microscope to visualize the arterial organization within the bone marrow near the metaphysis region (i) and the mid-shaft diaphysis region (j). k Nerves and arteries densities at different areas within the marrow space were quantified and normalized. Sampling locations are indicated with dotted lines in d, i, and j. SHG, second harmonic generation signal. All values are mean ± s.d., Statistical significance (*P < 0.05; **P < 0.01) was assessed by one-way ANOVA
