Scanning transmission electron microscopy methods for the analysis of nanoparticles

Here we review the scanning transmission electron microscopy (STEM) characterization technique and STEM imaging methods. We describe applications of STEM for studying inorganic nanoparticles, and other uses of STEM in biological and health sciences and discuss how to interpret STEM results. The STEM imaging mode has certain benefits compared with the broad-beam illumination mode; the main advantage is the collection of the information about the specimen using a high angular annular dark field (HAADF) detector, in which the images registered have different levels of contrast related to the chemical composition of the sample. Another advantage of its use in the analysis of biological samples is its contrast for thick stained sections, since HAADF images of samples with thickness of 100-120 nm have notoriously better contrast than those obtained by other techniques. Combining the HAADF-STEM imaging with the new aberration correction era, the STEM technique reaches a direct way to imaging the atomistic structure and composition of nanostructures at a sub-angstrom resolution. Thus, alloying in metallic nanoparticles is directly resolved at atomic scale by the HAADF-STEM imaging, and the comparison of the STEM images with results from simulations gives a very powerful way of analysis of structure and composition. The use of X-ray energy dispersive spectroscopy attached to the electron microscope for STEM mode is also described. In issues where characterization at the atomic scale of the interaction between metallic nanoparticles and biological systems is needed, all the associated techniques to STEM become powerful tools for the best understanding on how to use these particles in biomedical applications.