Fig. 2.

Cycle of SBF SEM imaging in the prototype apparatus by Leighton and Kuzirian. In a high-vacuum SEM chamber sections were cut from a resin block using a tungsten-coated glass knife, then the block face was etched with oxygen plasma, improving visualisation of tissue and cellular components. The block face was sputter-coated with gold to render it conductive and allow charge-free imaging before a secondary electron image was recorded and the cycle begun again. The best cycle time for sectioning, etching, coating and imaging was 10 min. Reproduced with the permission of Alan Kuzirian.