The top–bottom effect of a tilted thick specimen and its influence on electron tomography

We present the top–bottom effect (TBE) of a 5 µm thick amorphous specimen in a 3 MV ultrahigh voltage electron microscope (ultra-HVEM) and its influence on the quality of electron tomography (ET). The 40-nm gold particles on the top surface of the specimen tilted at different angles have been observed to be of poorer image quality than those on the symmetrical bottom surface obtained by tilting and turning over the specimen. The point spread function of the gold-particle images was calculated using the increment Wiener filter and the image-quality variation was then evaluated. The TBE is shown to become more remarkable with the increase of the effective thickness of the tilted specimen or the decrease of the magnification of the ultra-HVEM. The ET simulation indicates that the TBE may generate ∼4% distortion in the radial direction of a reconstructed sphere model.