โ Scribed by Turner, J. N. ;Rieder, C. L. ;Collins, D. N. ;Chang, B. B.
No coin nor oath required. For personal study only.
Optimal imaging of complex structures requires proper alignment relative to the optic axis of the electron microscope. This is especially important for high-voltage and intermediatevoltage microscopes, which form an in-focus image throughout the entire thickness of the object. As a result, structures at different specimen heights form overlapping and confused images that severely curtail the usefulness of these instruments.
The work described here provides a generalized, flexible method for optimizing specimen orientation and eliminating or limiting image overlap by means of a commonly used double-tilt stage. Analysis of the motion about the two axes provides accurate tilting for any azimuthal direction whether or not it corresponds to a mechanical axis of the stage. An object can be positioned to minimize image overlap, to record stereopairs for any parallax axis, and to record three-dimensional data sets by the conical collection geometry.
Images of muscle paracrystals are shown after tilting about an axis perpendicular to a symmetry direction. The tilted image displays higher-order symmetry, which is altered by changes of one degree. Precision double-tilting for optimizing stereopairs is shown for a desmosome recorded using different parallax axes and pretilts. A tomographic conical data-collection scheme is demonstrated by imaging a microtubule axoneme for a specific cone half-angle and arbitrary azimuthal angles.
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