Abstract
Abstract High quality imaging of the macromolecular structure of biological samples can be obtained when combining high resolution SEM with advanced thin metal film coating techniques. A thin layer of fine grain metal film is often necessary to enrich the secondary electron (SE) signal generation and to enhance the contrast of features of interest, because the yield of SE signal from biological samples is low. Secondary electron signals, types SE-I and SE-II, are generated by primary beam at the impact point or by backscattered electrons (BSE) dislocated from the impact point, respectively (Peters, 1982; Joy, 1984). The SE-II signal yield depends on the amount of BSE signal and the collection ratio of SE-I/SE-II components regulates the surface topographic contrast. Therefore, the low atomic number metals with low backscattering coefficients should be considered for high resolution SEM coating. Chromium, which has both the features of low atomic number (Z=24) and sufficient SE signal yield, is an appropriate choice and was first used by Peters (1982) for high resolution SEM imaging.
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