Institute of Fundamental Technological Research

Using high resolution focused ion beam scanning electron microscopy (FIB-SEM) we study the details of cell-nanostructure interactions using serial block face imaging. 3T3 Fibroblast cellular monolayers are cultured on flat glass as a control surface and on two types of nanostructured scaffold substrates made from silicon black (Nanograss) with low- and high nanowire density. After culturing for 72 hours the cells were fixed, heavy metal stained, embedded in resin, and processed with FIB-SEM block face imaging without removing the substrate. The sample preparation procedure, image acquisition and image post-processing were specifically optimised for cellular monolayers cultured on nanostructured substrates. Cells display a wide range of interactions with the nanostructures depending on the surface morphology, but also greatly varying from one cell to another on the same substrate, illustrating a wide phenotypic variability. Depending on the substrate and cell, we observe that cells could for instance: break the nanowires and engulf them, flatten the nanowires or simply reside on top of them. Given the complexity of interactions, we have categorised our observations and created an overview map. The results demonstrate that detailed nanoscale resolution images are required to begin understanding the wide variety of individual cells’ interactions with a structured substrate. The map will provide a framework for light microscopy studies of such interactions indicating what modes of interactions must be considered.

Nanowires, Thin films, Glass, Scanning electron microscopy, Transmission electron microscopy, Cell membranes, Fibroblasts, Nanomaterials

We present here a proof of concept for a novel fabrication method of vertically aligned carbon nanotube forests, utilizing black silicon nanograss (a forest of silicon nanometer-sized spikes created with reactive ion etching) coated with titanium tungsten diffusion barrier as a template. The method allows maskless definition of carbon nanotube forests with control of their density, nanotube diameter and height. Four nanograss reactive ion etching recipes are investigated and their wafer-to-wafer repeatability, wafer uniformity, and density control is discussed. Evaluation of carbon nanotube forests grown on the nanograss substrates is presented with discussion of their morphology, diameter distribution, and catalyst thickness influence.

Carbon nanotubes, Black silicon, Nanograss, Maskless, Catalyst, Titanium tungsten

Engineered nanomaterials (ENMs) are increasingly being used in the food industry. In order to assess the efficacy and the risks of these materials, it is essential to have access to methods that not only detect the nanomaterials, but also provide information on the characteristics of the materials (e.g., size and shape).

This review presents an overview of electron microscopy (EM)-based methods that have been, or have the potential to be, applied to imaging ENMs in foodstuffs. We provide an overview of approaches to sample preparation, including drying, chemical treatment, fixation and cryogenic methods. We then describe standard and non-standard EM-based approaches that are available for imaging prepared samples. Finally, we present a strategy for selecting the most appropriate method for a particular foodstuff.

Cryo-electron microscopy, Detection, Environmental electron microscopy, Foodstuff, Imaging, Liquid electron microscopy, Nanoparticle, Sample preparation, Scanning electron microscopy (SEM), Transmission electron microscopy (TEM)