Fig. 5

Correlated SEM and NanoSIMS inorganic ion images for room-temperature and cryogenic workflows. A SEM image of a section of conventionally prepared tissue of Green Hydra and corresponding NanoSIMS maps of the distributions of Na (B), Mg (C), K (D), and Ca (E). F Cryo-SEM image after a fully cryogenic sample preparation of tissue of Green Hydra and corresponding CryoNanoSIMS maps of the distributions of Na (G), Mg (H), K (I), and Ca (J). The images are from similar regions in the gastric region of Green Hydra animals that were exposed to identical incubation conditions (cf. Methods), permitting a direct comparison between the two analytical approaches. Note the dramatic loss of inorganic ions (especially Mg, K, and Ca) in conventional sample preparation due to dehydration and resin infiltration of the samples. Elemental maps are shown in a logarithmic color scale. These elemental maps are drift-corrected accumulations of 15 sequential images each consisting of 256 by 256 pixels with a pixel dwelling time of 5 ms; typical analysis time for one image as displayed is about 90 min. Scale bars are 5 µm. C, Chlorella sp. algae; EV, epidermal vacuole; E, epidermis; GVC, gastrovascular cavity; GV, gastrodermal vacuole; G, gastrodermis; M, mesoglea