Open Access Short report

Intracellular imaging of nanoparticles: Is it an elemental mistake to believe what you see?

Christina Brandenberger1*, Martin JD Clift12, Dimitri Vanhecke13, Christian Mühlfeld14, Vicki Stone2, Peter Gehr1 and Barbara Rothen-Rutishauser1

Author Affiliations

1 Institute of Anatomy, University of Bern, Baltzerstrasse 2, CH-3000, Bern 9, Switzerland

2 Centre for Nano Safety, School of Life Sciences, Edinburgh Napier University, Merchiston Campus, 10 Colinton Road, Edinburgh, EH10 5DT, UK

3 Max Planck Institute of Biochemistry, Department of Molecular Structural Biology, Martinsried, Germany

4 Institute of Anatomy and Cell Biology, University of Giessen, Giessen, Germany

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Particle and Fibre Toxicology 2010, 7:15  doi:10.1186/1743-8977-7-15

Published: 3 June 2010


In order to understand how nanoparticles (NPs <100 nm) interact with cellular systems, potentially causing adverse effects, it is important to be able to detect and localize them within cells. Due to the small size of NPs, transmission electron microscopy (TEM) is an appropriate technique to use for visualizing NPs inside cells, since light microscopy fails to resolve them at a single particle level. However, the presence of other cellular and non-cellular nano-sized structures in TEM cell samples, which may resemble NPs in size, morphology and electron density, can obstruct the precise intracellular identification of NPs. Therefore, elemental analysis is recommended to confirm the presence of NPs inside the cell. The present study highlights the necessity to perform elemental analysis, specifically energy filtering TEM, to confirm intracellular NP localization using the example of quantum dots (QDs). Recently, QDs have gained increased attention due to their fluorescent characteristics, and possible applications for biomedical imaging have been suggested. Nevertheless, potential adverse effects cannot be excluded and some studies point to a correlation between intracellular particle localization and toxic effects.

J774.A1 murine macrophage-like cells were exposed to NH2 polyethylene (PEG) QDs and elemental co-localization analysis of two elements present in the QDs (sulfur and cadmium) was performed on putative intracellular QDs with electron spectroscopic imaging (ESI). Both elements were shown on a single particle level and QDs were confirmed to be located inside intracellular vesicles. Nevertheless, ESI analysis showed that not all nano-sized structures, initially identified as QDs, were confirmed. This observation emphasizes the necessity to perform elemental analysis when investigating intracellular NP localization using TEM.