Zero‐loss energy filtering under low‐dose conditions using a post‐column energy filter

Electron cryomicroscopy combined with energy filtering can be performed under low-dose conditions using a post-column energy filter. The microscope combined with the filter is set up such that it can be used with similar ease as a conventional microscope, the main difference being that all filter and microscope control is performed through a central computer and images are recorded with a cooled slow-scan CCD camera. The microscope can also still be used for regular imaging on film as without the filter. Owing to the 18 times post-magnification of the filter, the microscope normally has to be operated at a small magnification, e.g. 3000×, and the beam has to be contracted to a small spot, e.g. 5 mm, in the plane of the microscope viewing screen. Computer control allows one to perform a variety of tasks automatically, such as autofocusing, thickness measurements, most-probable-loss imaging, CCD spot-scanning and tomography. The gain in contrast due to zero-loss energy filtering is analysed using visual inspection, power spectra and Fourier ring correlation. The thickness range for ice-embedded specimens in which a filter at 120 kV is most useful appears to be between 100 and 300 nm.