The different binding modes of Gx-DiMan with DC-SIGN/R were further verified by "cryo-snapshot" TEM imaging, which has been shown to be able to capture nanoparticle in their native dispersion states in solution. This was achieved by rapid plunge-freezing of the sample into liquid ethane, followed by drying under vacuum and finally loading on the TEM grids for TEM imaging. Here, G27-DiMan was employed in the investigation. G27-DiMan was first mixed with DC-SIGN or DC-SIGNR at a PGR of 80 and incubated for 40 mins, then they were plunge-frozen for sample preparation and finally applied for TEM imaging. The resulting TEM images (Figs. ) clearly revealed that binding of DC-SIGN with G27-DiMan gave completely isolated single particles, whereas binding of DC-SIGNR produced large-scale clustered GNPs. These results fully agreed with their Dh sizes described in the previous section. The combined TEM and DLS data thus reaffirmed the distinct modes for DC-SIGN/R in binding to G27-DiMan, where DC-SIGN binds simultaneously with all four binding sites to one G27-DiMan to form small, isolated GNP/protein core/shell particles, while DC-SIGNR crosslinks with different G27-DiMan particles to form large scale GNP-protein assemblies. These results completely agree with those observed previously between DC-SIGN/R and G5-DiMan or (4 nm) QD-DiMan. Therefore, increasing the Gx-DiMan scaffold size from 5 to 27 nm did not alter their binding mode with DC-SIGN/R in the solution, although it did significantly enhance their MLGI affinities.