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Fig. 11 Flow chart describing the automation process for a single tilt series reconstruction.
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.
In Fig. 4, the front (XY), top (XZ) and side (YZ) views of a Mimivirus reconstruction are displayed for a single tilt (N = 1, Fig. 4A top row) and a 16-tilt (N = 16, Fig. 4A bottom row) series; the uppercase letters X, Y, and Z are used to specify the coordinates of a point in the specimen, Z being in the normal direction to the specimen slab.
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.
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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.
For each experimental condition (Table 1), we collected three conical series using a Gatan 650 Single Tilt Rotating Holder in a FEI Tecnai 12 electron microscope at 120 KV.
Even a single engine tilt adjustment at the beginning of deflection operations yields more deflection than a fixed preset tilt.
Automated data acquisition of the single tilt series through an angular range of −69° to +69° with 1° increments was performed using a field emission gun electron microscope operating at 200 kV (Tecnai F20; FEI Company, Eindhoven, The Netherlands).
Automated data acquisition of the single tilt series through an angular range of −65° to +65° with 1° increments was performed using an electron microscope equipped with a field emission gun and operating at 200 kV (Tecnai F20, FEI Company, Eindhoven, The Netherlands) and Xplore 3D.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com