We describe techniques used to characterize hierarchically structured synthetic and biological materials. These techniques decipher the structures through the dimensional spectrum from the molecular and atomic scales (10.−10 m) to the macro scales (10−3 m) where the overall shape of the material emerges. Techniques used to image surfaces and internal structures can be categorized according to their wavelength and thus spatial resolution as light, x-ray, and electron microscopy. We also discuss newly emerging microscopy techniques that image surfaces at the atomic level without a focusing lens, such as scanning tunneling and atomic force microscopies, with the aid of field ion microscope and scanned probe microscopes. Transmission electron microscopy (TEM), a unique tool for multipurpose imaging, provides structural information through direct imaging, diffraction, and spectroscopic analysis. We illustrate the major TEM techniques used to analyze structural hierarchy with examples of synthetic and biological materials. We also describe light optical microscopy and scanning probe microscopy techniques, which cover the opposite ranges of the dimensional spectrum at the micrometer and subangstrom levels.