Abstract
Cellular morphology is inherently three-dimensional. However, most histological techniques for tissue analysis focus on extracting information from two-dimensional slices of fixed samples or dissociated cells. These techniques result in a significant loss of the three-dimensional information of the tissue, including true cell volume, orientation, and whole cell shape. This unit discusses various options for three-dimensional imaging, provides a protocol for performing post-processing reconstruction based on serial slicing, and discusses the current advantages and limitations of the three-dimensional approach to quantitative tissue analysis. The focus of this protocol is on cardiac tissue, but the techniques can be applied to any solid tissue.