3D Local in vivo Environment (LivE) imaging for single cell protein analysis of bone tissue: LivE imaging is a novel methodological platform for linking individual expression profiles of cells within bone tissues to their local remodeling and mechanical environments

The molecular processes behind pathological bone remodelling seen in diseases such as osteoporosis are unclear. However, a recently developed methodological platform known as Local in vivo Environment (LivE) imaging has been used to link cellular expression data to the local remodelling and mechanical environment in 2D sections of bone tissue. The method therefore can be used to give insight into which proteins are important for pathological bone remodelling. However, the cells within bone tissue exist as a 3D network. Therefore extension of LivE to accommodate 3D data may provide additional physiologically relevant information that is not possible to determine using 2D analysis alone. This will have implications for the further understanding of the cellular basis that underlies bone diseases such as osteoporosis. Here the LivE imaging technique is expanded to incorporate data from cells in a three dimensional manner via a serial sectioning technique. The methodological steps involved in the LivE imaging approach are defined and the optimisation steps performed are explained in detail.