Development of methodology for low exposure, high resolution electron microscopy of biological specimens.
Specimen damage resulting from inelastic scattering is one of the factors that limits high-resolution electron microscopy of biological specimens. We have, therefore, sought to develop a method to record images of periodic objects at a reduced electron exposure in order to preserve high-resolution structural detail. The resulting image will tend increasingly to be a statistically noisy one, as the electron exposure is reduced to lower and lower values. Construction of a statistically defined image from such data is possible by spatial averaging of the electron signals from a large number of identical unit cells. In this paper, we have first investigated the theory pertaining to the attainable resolution as a function of the electron exposure, the magnification, and several other relevant parameters. In addition, we report experimental results obtained with a commercial image intensifier and with nuclear track photographic emulsion, both of which are highly sensitive recording devices. Usable images can be recorded and processed at exposures in the image plane as low as 10(-3) electron/micron2 (1.6 x 10(-14) coulomb/cm2).
D004583:Electrons; D008854:Microscopy, Electron / Q000295:instrumentation / Q000379:methods*; D008962:Models, Theoretical; D012542:Scattering, Radiation