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The tilt series is collected using linear tilt scheme continuously from -70° to +70°with tilt increments of 2°.
In general, datasets with limited tilt range tend to result in reconstructions with artefacts that are harder to correct than those related to large tilt increments.
Series up to 16-fold rotation axis and 1° tilt increments between −60° and +60° were acquired for this study; in this extreme scenario (with around 2000 projection views), the data acquisition step was completed within a single day.
Although automated low-dose ET acquisition packages have been developed and algorithms that tackle datasets with large tilt increments have been successfully implemented, I-DS techniques based on recording a subset of pixels have not yet, to our knowledge, been tested on ET datasets.
K2 images were recorded at 2° tilt increments, with −5 μm defocus, at 11,800× and 14,500× magnifications (pixel sizes of 4.2 Å and 3.4 Å).
For each reconstruction, a series of images at regular tilt increments was collected with a JEOL 4000EX intermediate-voltage electron microscope operated at 400 kV.
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The nominal tilt range was ±66°, tilt increment was 1.5° or 2°, and cumulative dose was 100 140 electrons/Å.
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.
Tilt increment was 2°.
T-DS techniques rely on either reducing the tilt range or increasing the tilt increment.
Video of an undersampled tilt series, corresponding to 60% I-DS and 2° tilt increment.
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