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Serial Block-Face Imaging by Field Emission Scanning Electron Microscopy  

Kim, Ki-Woo (School of Ecology and Environmental System, Kyungpook National University)
Publication Information
Applied Microscopy / v.41, no.3, 2011 , pp. 147-154 More about this Journal
Abstract
Backscattered electrons (BSE) are generated at the impact of the primary electron beam on the specimen. BSE imaging provides the compositional contrast to resolve chemical features of sectioned block-face. A focused ion beam (FIB) column can be combined with a field emission scanning electron microscope (FESEM) to ensure a dual (or cross)-beam system (FIB-FESEM). Due to the milling of the specimen material by 10 to 100 nm with the gallium ion beam, FIB-FESEM allows the serial block-face (SBF) imaging of plastic-embedded specimens with high z-axis resolution. After contrast inversion, BSE images are similar to transmitted electron images by transmission electron microscopy. As another means of SBF imaging, a specialized ultramirotome has been incorporated into the specimen chamber of FESEM ($3View^{(R)}$). Internal structures of plastic-embedded specimens can be serially revealed and analyzed by $3View^{(R)}$ with a large field of view to facilitate three-dimensional reconstruction. These two SBF approaches by FESEM can be employed to unravel spatial association of (sub)cellular entities for a comprehensive understanding of complex biological systems.
Keywords
3View; Focused ion beam; Serial block-face; Tomography;
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Times Cited By KSCI : 1  (Citation Analysis)
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