Because of the importance of diatoms and the lack of information about their third dimension, a new method for the 3D reconstruction is explored, based on digital image correlation of several scanning electron microscope images. The accuracy of the method to reconstruct both centric and pennate (symmetrical and asymmetrical) diatoms was shown, independently of valve size and shape, and considering not only the general frustule morphology but also the intricate ornamentation. Several measurements were obtained, such as of the surface and projected areas and the valve volume. These results were validated by focused ion beam transverse cross section of one valve, and the quantitative results were compared with geometric models commonly applied. Furthermore, direct volume calculations based on 3D reconstructions have several advantages, such as it has higher taxonomic accuracy than the methods based on light microscopy; it solves the problems of the light halos; it allows the precise measurement of all linear dimensions, including the often neglected third dimension; natural samples can be measured directly; and it provides an exact estimate of cell volume, independently of its shape or alterations due to life cycle stage. This approach provides therefore a simple way to measure the morphological features of diatoms, even at a nanoscale, and can be applied to other microorganisms commonly illustrated by means of scanning electron microscopy.