Pt/Ir/C, a new, powerful coating material for High-Resolution SEM
In-lens field emission SEM allows to image specimen surfaces with subnanometer resolution by collecting type I secondary electrons, with an incident electron probe size of abut 0.5 nm in diameter. To achieve such high topographic resolution on biological specimens adequate structural preservation and high resolving thin continuous coating films are necessary.Careful freeze-drying (“maintaining hydration shells”) followed by heavy metal shadowing at -250°C under ultra high vacuum conditions (UHV) allows to extract surface features ≤2 nm with TEM. TEM-shadowing films (Pt/C, Ta/W) are stabilized with a C-backing layer. Such a C-coat would blurr fine details when investigated with the SEM.At the right composition (C-content 25-40%, metal content 75-60%) only about 1.5 nm thick Pt/Ir/C-films remain three-dimensionally stable when transferred to atmospheric conditions after freeze-drying samples with macromolecular dimensions. Pt/Ir/C is made by evaporating a Pt/Ir cylinder (diameter 1.5 mm, 70% Ir) inserted into a graphite rod (diameter 2 mm).