Extended Data Fig. 1: Comparison of grid adsorption orientation and signal delocalization for two chassis designs.
From: Molecular goniometers for single-particle cryo-electron microscopy of DNA-binding proteins
a, Representative negative-stain micrographs of chassis with 0.67 aspect ratio. b, Representative cryo-EM micrograph of chassis with 0.59 aspect ratio. Desired orientations assessed by manual counting. c, Top: Representative 2D class average of 0°-rotation goniometer from a. The chassis aperture exhibits a shadow that overlaps the DNA stage and BurrH protein. Bottom: Representative 2D class averages of BurrH, exhibiting artifacts that we hypothesized are due to delocalized origami signal in the 0.5–2.0 µm defocus range used for image acquisition. d, Top: 2D class of 0°-rotation goniometer from b designed to reduce shadow overlap. Bottom: Representative BurrH 2D class averages. Artifacts observed in c are significantly reduced. Scale bars: a, b: 100 nm, c, d: 25 nm (top), 10 nm (bottom).
