Application of the low vacuum scanning electron microscope to the study of biomaterials and mammalian cells

The use of the scanning electron microscope (SEM) in 'low' (reduced) vacuum (lvac) mode permits observation of specimens which have not been coated with a conductive material such as gold or carbon. We have evaluated the use of this mode of observation to the study of biomaterials using the bone-substitute material Interpore as an example. On this material, rat bone cells were visible in lvac mode only in cells traversing pores, when they were readily identified by their cell nuclei. Rat calvarial bone examined uncoated in lvac mode showed the bone structure clearly through the overlying layer of osteoblast cells, which were subsequently revealed by gold coating. Immunogold labelling of alkaline phosphatase was imaged in lvac mode, following silver enhancement and carbon coating. These studies demonstrate the complementary use of the lvac and high vacuum (hvac) SEM to study material composition, the behaviour of mammalian cells on biomaterials and the potential use of lvac SEM to study mineralized tissues without removal of overlying soft tissue.