Figure 4.

The effect of decalcification on CGRP, NGF, TrkA, and p75 immunostaining in the femur of the young mouse. These confocal images were taken from young bones that had been fixed and then placed for 14 days in either PBS solution (pH = 7.4, 4℃) (A, C, E, G) or a decalcification solution consisting of PBS + EDTA (pH = 7.4, 4℃) (B, D, F, H). After 14 days, both the decalcified and non-decalcified femurs were placed in a 30% sucrose solution for two days, frozen sections cut, and the sections then immunostained for CGRP, TrkA, p75, or NGF. Note that robust immunohistochemical staining is observed for CGRP, TrkA, p75, and NGF in bones that were not decalcified (A, C, E, G). In contrast, in bones that had been decalcified, CGRP showed no significant loss of signal following decalcification (B), whereas TrkA (D), p75 (F), and NGF (H) all showed significant loss of immunohistochemical signal. All images presented here were obtained in the bone marrow. z-stack = 60 µm, line bar = 30 µm.