Micrographs were collected on a Titan Krios G1 electron microscope (FEI) operated at 300 kV, equipped with a GIF Quantum LS imaging filter (Gatan) and a K2 Summit detector (Gatan) operating in counting mode, using SerialEM. A total of 6,336 movies were acquired at a nominal magnification of 165,000×, corresponding to a pixel size of 0.82 Å, at defocus values ranging from 0.8–2.0 μm. Each movie was collected over a 9 s exposure, fractionated into 40 frames, with a total dose of approximately 51 e−/Å2. Images were acquired using beam-image shift to record 4 images per hole, except for initial test images.
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