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We did not try to get 2D averages from the cryoEM images because the signal from a single tilt series image (at low magnification and low dose compared to single particle cryo-EM images) is too weak to distinguish small features.
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Fig. 11 Flow chart describing the automation process for a single tilt series reconstruction.
A single tilt series contains up to 90 individual diffraction patterns collected from a single crystal with tilt angle increment of 0.1 1° and a total accumulated electron dose less than 10 electrons per angstrom squared.
It appears the signal amplitude of a single tilt (N = 1) reconstruction is higher compared to a 16-tilt reconstruction.
Tilt series were collected along a single tilt axis (Fig. 4B) with attentions paid to avoid the electron irradiation damage.
The Cu grids were transferred to the microscope using a single tilt holder.
A single tilt specimen holder was used, and images and diffraction patterns were recorded on Kodak film.
To obtain an optimal coverage of the complexes during a single-tilt tomographic image acquisition, the long axis of the complexes had to run either parallel to the tilt axis or parallel to the beam axis of the microscope.
(A and B ) Two examples of diffraction patterns obtained from a single crystal at tilt angles of 0° and 20° respectively.
Even a single engine tilt adjustment at the beginning of deflection operations yields more deflection than a fixed preset tilt.
A second pass might be unnecessary for single tilt series.
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