Imaging colloidal gold labels in LVSEM

Abstract

AbstractOn biological samples, the topographic imaging capabilities of the new generation of scanning electron microscopes (SEM) (those having both field‐emission guns and low aberration lenses) rival those of the replica techniques. In addition, they permit the localization of specific molecules on the sample surface using one of several labeling techniques utilizing heavy metal colloids. Normally, colloidal gold can be detected in the SEM both by the secondary electron signal (shape) and by the backscattered electron signal (BSE, Z‐contrast). The new instruments seem to produce their best topographic images using low‐beam voltage (1–5 kV) where topographic contrast is higher and the required thickness of the metal coating is less (Haggis and Pawley 1988, Ris and Pawley 1988). Although the detection of backscattered electrons is more difficult at low‐beam voltage, we are able to show here that the secondary electron (SE) signal produced with a 2–5‐kV beam permits the unambiguous detection of gold particles as small as 5 nm on carbon‐coated specimens while a 1‐kV beam produces a high‐quality topographic image of the same sample.