Abstract
Serial block-face scanning electron microscopy (SBF-SEM) is an advanced 3-D electron microscopy for investigating structures of large volume specimens such as a whole cell or a piece of tissues at a resolution better than a few tens of nanometers. In this method, block surface of a resin embedded specimen is trimmed by a diamond knife attached to an in-chamber ultramicrotome, and the newly exposed surface is imaged by SEM. The sectioning and imaging can be repeated automatically to get serial block-face images of the specimen. The resultant 3-D structure is reconstructed from the serial block-face images after individual image alignment. This review shows the principle and performance of SBF-SEM, and introduces some recent applications using SBF-SEM. For example, morphological changes in mitochondria regulated by Cdc48p/p97 ATPase were recently examined by SBF-SEM. The Cdc48p is a highly conserved cytosolic AAA chaperone that involved in a wide range of cellular processes including the regulation of the mitochondrial morphology. The results demonstrate that SBF-SEM has considerable advantages in quantitative morphological analyses on organelles and intracellular structures in the whole cell.
