Methods for specimen thickness determination in electron microscopy. II. Changes in thickness with dose

The electron diffraction patterns of tilted thin crystals were used to determine the unit cell size in the direction normal to the supporting film. The method revealed a considerable dose-dependent thinning or shrinkage. Using a variety of specimens and stains, we found that this amounted to a 50% reduction in volume and could be attributable to two causes. Firstly, the specimen is held to the supporting film so that volume changes can only occur through changes in thickness. Secondly, the decrease in volume is associated with a dose-induced mass loss which is greatly suppressed at liquid nitrogen temperatures.