Journal article
Mapping the Complex Morphology of Cell Interactions with Nanowire Substrates Using FIB-SEM
Molecular Windows, Department of Micro- and Nanotechnology, Technical University of Denmark1
Department of Micro- and Nanotechnology, Technical University of Denmark2
Materials and Surface Engineering, Department of Mechanical Engineering, Technical University of Denmark3
Department of Mechanical Engineering, Technical University of Denmark4
Nanoprobes, Department of Micro- and Nanotechnology, Technical University of Denmark5
Fluidic Array Systems and Technology, Department of Micro- and Nanotechnology, Technical University of Denmark6
University of Copenhagen7
Center for Electron Nanoscopy, Technical University of Denmark8
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.
Language: | English |
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Publisher: | Public Library of Science |
Year: | 2013 |
Pages: | e53307 |
ISSN: | 19326203 , 15537358 and 1553734x |
Types: | Journal article |
DOI: | 10.1371/journal.pone.0053307 |
ORCIDs: | Jensen, Mikkel Ravn Boye and Mølhave, Kristian |