Overlay of confocal images of whole mount preparations onto μCT images of bone showing sprouting and neuroma formation by sensory nerve fibers. Confocal images of whole mount preparations of the femoral periosteum from sham (A) or sarcoma + vehicle mice (B) immunostained with calcitonin gene-related peptide (CGRP) and overlayed onto uCT images of the bones from which they were obtained. Confocal images of periosteum (approximately 70 μm in thickness) were acquired from whole mount preparations and projected from 280 optical sections at 0.25 μm intervals with a 40x objective. Z-stack images of whole mount preparations from four sham or sarcoma + vehicle mice were acquired, tiled, and overlaid to scale on a three-dimensional micro-CT image of the sham femur (A) and sarcoma + vehicle femur (B), respectively using AMARA software. Note that the tumor-injected femur (B) has severe cortical bone deterioration and a pathological reorganization of CGRP nerve fibers (in red) compared to the sham femur (A).