Fig. 9.

Effects of varying spot size and accelerating voltage on SBF SEM image quality. Images were taken using SBF SEM at a pixel size of 3.8 nm, 4k × 4k image size, a dwell time of 4 μs and a chamber pressure of 60.0 Pa. Images captured at lower accelerating voltage showed lower image contrast, while increased accelerating voltage and larger spot size led to damage on the block surface and charging, shown by dark patches in the image and lost detail within the cell. The image on the left shows neither charging nor surface damage and exhibits adequate image contrast to distinguish details of the cell ultrastructure. All scale bars = 2 μm.