Fig. 8.

Beam/sample interactions during SBF SEM imaging. (A) The incident electron beam (black arrow) interacts with the sample and BSEs (blue arrows) are detected. The block moves upwards (white arrow) and the knife moves horizontally (double-headed arrow) to remove a slice of tissue before the cycle restarts. (B) Increasing accelerating voltages (grey arrows) lead to a greater depth of electron penetration and associated tissue damage. Slice thickness (dotted lines) should be greater than the penetration depth of the beam in order to remove resin which has been affected by the beam and thus, to avoid surface damage in the subsequent image. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)