Density-Dependent Color Scanning Electron Microscopy (DDC-SEM) for calcified tissue and pathological calcification.

Scanning electron microscopy (SEM) is widely used for materials characterization. It has also been successfully applied to the imaging of biological samples, providing invaluable insights into the topography, morphology and composition of biological structures, including pathological minerals, in diseases affecting cardiovascular, kidney and ocular tissues. Here we provide a comprehensive and detailed guide on how to use colored SEM to aid the visualization and characterization of pathological calcification, and identify the effects of different sample preparation protocols for the visualisation of these minerals.