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KMID : 0381920110410030147
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Korean Journal of Microscopy
2011 Volume.41 No. 3 p.147 ~ p.154
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Serial Block-Face Imaging by Field Emission Scanning Electron Microscopy
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Kim Ki-Woo
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Abstract
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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®). Internal structures of plastic-embedded specimens can be serially revealed and analyzed by 3View® with a large field of view tofacilitate 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.
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KEYWORD
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3View, Focused ion beam, Serial block-face, Tomography
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