Abstract
Computer control of a transmission electron microscope (TEM) and digital recording of images have shown to be useful tools in recording 2D data sets of macromolecules and larger cellular structures with accurately defined imaging conditions.3D structure of individual molecules can be obtained by electron tomography. The method requires that projection data are collected over a large range of specimen tilts. With automated electron tomography images are recorded in digital format, and lateral specimen displacement together with focus changes are automatically compensated for. Typically, with our experimental set-up, we collect 75 projections of negatively stained preparations over ±75° and 30 projections of ice-embedded molecules over ±60° with a total dose of 20-50 e/Å2.By using the spot-scan imaging mode beam-induced movements of ice-embedded specimens are greatly reduced, thus largely avoiding the loss of resolution associated with specimen instability. With CCD spot-scan imaging the size of the spot is matched to the CCD chip (Fig. 1).
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Schedule a callMore From: Proceedings, annual meeting, Electron Microscopy Society of America
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