Hypothesis for a mechanism of beam-induced motion in cryo-electron microscopy

Abstract

Estimates of heat-transfer rates during plunge-cooling and the patterns of ice observed in cryo-EM samples indicate that the grid bars cool much more slowly than do the support foil and sample near the middle of the grid openings. The resulting transient temperature differences generate transient tensile stresses in the support foil. Most of this foil stress develops while the sample is liquid and cooling toward its glass transition Tg, and so does not generate tensile sample stress. As the grid bars continue cooling towards the cryogen temperature and contracting, the tensile stress in the foil is released, placing the sample in compressive stress. Radiation-induced creep in the presence of this compressive stress should generate a doming of the sample in the foil openings, as is observed experimentally. Crude estimates of the magnitude of the doming that may be generated by this mechanism are consistent with observation. Several approaches to reducing beam-induced motion are discussed.

Keywords

• cryo-em
• beam-induced motion
• radiation damage