Extended Data Fig. 5: PACE-tomo facilitates data collection in challenging areas.

a. Shown is a 0.66 nm thick slice through one of the 21 cryo electron tomograms (acquisition area 3) collected on the lamella shown in Fig. 4b. The subtomogram average was mapped into the tomogram using the refined coordinates and orientations. Membrane associated and cytoplasmic ribosomes were coloured red and blue, respectively. Scale bar: 100 nm. b. Shown is a 0.66 nm thick slice through one of 7 cryo electron tomograms collected on the bottom lamella shown in c. The subtomogram average of actin was mapped into the tomogram using the refined coordinates and orientations. Scale bar: 100 nm. c. Overview montages of two (out of 41) cryoFIB-milled lamellae indicating the positions at which tilt series were collected. Red frame indicates the position that was used for the tracking tilt series. White arrowheads mark targets that would not have been accessible by conventional cryoET collection schemes. Collection times were 23 minutes for 6 tilt series (top) and 30 minutes for 7 tilt series (bottom, used in dataset 5).