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Electron tomography has become a commonly used tool to investigate thethree-dimensional (3D) structure of nanomaterials, including colloidalnanoparticle assemblies. However, electron microscopy is typically carried outunder high vacuum conditions. Therefore, pre-treatment sample preparation isneeded for assemblies obtained by (wet) colloid chemistry methods, includingsolvent evaporation and deposition on a solid TEM support. As a result of thisprocedure, changes are consistently imposed on the actual nanoparticleorganization. Therefore, we propose herein the application of electrontomography of nanoparticle assemblies while in their original colloidal liquidenvironment. To address the challenges related to electron tomography inliquid, we devised a method that combines fast data acquisition in a commercialliquid-TEM cell, with a dedicated alignment and reconstruction workflow. Wepresent the application of this method to two different systems, whichexemplify the difference between conventional and liquid tomography, dependingon the nature of the protecting ligands. 3D reconstructions of assembliescomprising polystyrene-capped Au nanoparticles encapsulated in polymeric shellsrevealed less compact and more distorted configurations for experimentsperformed in a liquid medium compared to their dried counterparts. On the otherhand, quantitative analysis of the surface-to-surface distance ofself-assembled Au nanorods in water agrees with previously reported dimensionsof the ligand layers surrounding the nanorods, which are in much closer contactwhen in similar but dried assemblies. This study, therefore, emphasizes theimportance of developing high-resolution characterization tools that preservethe native environment of colloidal nanostructures.

Liquid phase fast electron tomography unravels the true 3D structure of colloidal assemblies