Fig. 2.

Image shift compensation can be used to minimize specimen shifts. A/B: Specimen shift throughout a FISE dose-symmetric 3° increment tilt series recorded without application of compensating image shifts (A) and with application of compensating image shifts (B) recorded on a Titan Krios [ST/K3]. Image shifts effectively compensate specimen shifts, especially at large tilt angles. C/D: A single calibration tilt series can be used to compensate specimen shifts for multiple grid squares. The schematic (C) shows squares of a Quantifoil Cu 200 R2/2 grid and refers to the relative positions of tilt series that were collected with (colored) and without (grey) compensating image shifts (D). Shown are specimen shifts perpendicular to the tilt axis (y-shift) within full tilt series. E: Y-shifts of 47 tilt series collected in less than four hours on eleven different squares (Fig. S3) using a single calibration tilt series and a Titan Krios [ST/K3]. Blue crosses mark tilt angles higher than ±51°. The arrowhead indicates the worst tilt series that is also shown in Movie S3. Tilt series are grouped by squares. No micrographs had to be discarded due to the loss of target.