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Abstract
Standard transmission electron microscopy nanoparticle sample preparation generally requires the complete removal of the suspending liquid. Drying often introduces artifacts, which can obscure the state of the dispersion prior to drying and preclude automated image analysis typically used to obtain number-weighted particle size distribution. Here we present a straightforward protocol for prevention of the onset of drying artifacts, thereby allowing the preservation of in-situ colloidal features of nanoparticles during TEM sample preparation. This is achieved by adding a suitable macromolecular agent to the suspension. Both research- and economically-relevant particles with high polydispersity and/or shape anisotropy are easily characterized following our approach (http://bsa.bionanomaterials.ch), which allows for rapid and quantitative classification in terms of dimensionality and size: features that are major targets of European Union recommendations and legislation.
Highlights
Standard transmission electron microscopy nanoparticle sample preparation generally requires the complete removal of the suspending liquid
We present a straightforward protocol for prevention of the onset of drying artifacts, thereby allowing the preservation of in-situ colloidal features of nanoparticles during transmission electron microscopes (TEM) sample preparation
We distinguished single particles from ex situ aggregates and always compared samples prepared in the standard manner to those prepared using our method, which consisted of mixing an aqueous solution of Bovine serum albumin (BSA) with the Au NPs, drop casting the suspension onto the TEM grid, and letting it dry under ambient conditions
Summary
Standard transmission electron microscopy nanoparticle sample preparation generally requires the complete removal of the suspending liquid. We present a straightforward protocol for prevention of the onset of drying artifacts, thereby allowing the preservation of in-situ colloidal features of nanoparticles during TEM sample preparation This is achieved by adding a suitable macromolecular agent to the suspension. Electron microscopy is a so-called counting method, which determines individual nanoparticle size and can be used to construct the required number-weighted size distributions This technique is frequently plagued with issues related to artifacts, statistical reliability and interpretation.[2] Drying steps, unavoidable during sample preparation, can result in non-uniform particle deposition and particle aggregation.[3] Characterizing commercially-relevant materials, which often have highly non-uniform sizes and shapes, is challenging. BSA is not necessarily the only possible choice, it was chosen as it is cheap, widely available and well-studied
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