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Figure 3
Phase-plate tomography of the perforin pre-pore complex. The 0° image from the tomogram (a), the central ten z sections averaged from the reconstructed tomogram (b) and an enlarged 20-z-section average from the reconstructed tomogram (c) of perforin pre-pores bound to liposomes collected with the phase plate. The tomogram was collected on Krios II at eBIC at 1.7 Å per pixel using the Quantum K2 detector in counting mode (∼5 electrons per pixel per second) with a 20 eV slit width and a nominal defocus of 300 nm to avoid going over focus. Tilt images were collected from −45 to 45° in 3° increments, giving a total dose of around 60 e Å−2. The tomogram was collected using the FEI TOMO package and was processed with MotionCorr and IMOD (Li et al., 2013BB21; Kremer et al., 1996BB19). The scale bars in (a) and (b) are 100 nm and the scale bar in (c) is 10 nm. The grids were prepared by N. Lukoyanova.

Journal logoSTRUCTURAL
ISSN: 2059-7983
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