title: Mapping the complex morphology of cell interactions with nanowire substrates using FIB-SEM.
abstract: 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.
authors: DTU Nanotech, Technical University of Denmark, Lyngby, Denmark.
authors: Rafał
authors: R
authors: Wierzbicki
authors: Carsten
authors: C
authors: Købler
authors: Mikkel R B
authors: MR
authors: Jensen
authors: Joanna
authors: J
authors: Lopacińska
authors: Michael S
authors: MS
authors: Schmidt
authors: Maciej
authors: M
authors: Skolimowski
authors: Fabien
authors: F
authors: Abeille
authors: Klaus
authors: K
authors: Qvortrup
authors: Kristian
authors: K
authors: Mølhave
mesh_terms: D000818:Animals; D002450:Cell Communication / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure*; D021621:Imaging, Three-Dimensional; D051379:Mice; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D041681:NIH 3T3 Cells; D053770:Nanowires / Q000648:ultrastructure; D012825:Silicon / Q000494:pharmacology
chemical_list: D012825:Silicon