Advantages of FIB/SEM Tomography in SEM-Based Three-Dimensional Reconstruction
Department of Anatomy, Kurume University School of Medicine
Abstract: The combination of focused ion-beam milling with scanning electron microscopy (SEM) backscattered imaging provides a versatile three-dimensional (3D) reconstruction method (FIB/SEM tomography) that can visualize the hierarchical structural organization of tissue, cell and organelle with a similar resolution in electron microscopy. The nanofabrication technology of FIB allows milling of various solid materials—including bone tissue, which is one of difficult material to cut off with mechanical methods—and following 3D reconstruction of the specimen at resolution as good as that of an electron tomography (>5 nm). Although the possible size of the 3D reconstruction is limited to tens of micrometers cubic, the survey specimen sizes are not limited. Thus, 3D reconstruction targets can be selected by an overview of a large specimen area. This procedure is useful for biomedical materials as well as clinical subjects and the correlative observation of light and electron microscopy. The method is thus indispensable for in imaging of cell and tissue architectures in their entirety at nanometer-range resolution.
Key words: FIB/SEM tomography, biomedical application, mitochondria, block face image