Coherent, non-planar illumination of a defocused specimen: consequences for transmission electron microscopy

Abstract

Summary The objective of this study has been to examine the imaging properties of transmission electron microscopes when coherent non-planar illumination is used in conjunction with defocused specimens. This situation is reminiscent of what is commonly the case in electron microscopic phase-contrast studies of biologically relevant macromolecules, using a field emission gun as a coherent electron source. For the sake of simplicity, the imaging system has been idealized as a thin lens with properties that can be described by the Fresnel approximation of the Huygens-Fresnel principle. The resulting expressions show that the system magnification has a defocus dependent factor, as do the contrast transfer functions. These factors are normally not taken into account in conventional derivations. The defocus dependent factor can be minimized by using planar illumination. The factor approaches infinity as the crossover moves closer to the specimen, and it is in the region close to the specimen that this factor is most significant. These results can have serious implications for high-resolution single-particle cryo electron microscopy as this technique often relies on combining data taken at a range of defocus values.