4.7 Article

Quantification of nanoparticle uptake by cells using an unbiased sampling method and electron microscopy

期刊

NANOMEDICINE
卷 6, 期 7, 页码 1189-1198

出版社

FUTURE MEDICINE LTD
DOI: 10.2217/NNM.11.70

关键词

3T3; cellular uptake; fractionator; gold nanoparticle; mouse fibroblast; nanoparticle; nanoparticle pathways; quantification; stereology; TEM

资金

  1. European Union [NMP4-CT-2006-033231, NMP4-SL-2008-214547]

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Aims: By randomly sampling a known fraction of a pellet of cultured cells, we have accurately estimated the mean number of 50 nm gold nanoparticles accumulated within a single cell. Cellular nanoparticle uptake was measured using a combination of stereological sampling techniques and transmission electron microscopy. Materials & Methods: Nanoparticles were counted individually and their intracellular location was recorded. Quantifying cell and nanoparticle number by analyzing a known fraction of the sample led to precise estimates of intracellular nanoparticle numbers and their spatial locations on an ultrastructural level. We propose a simple and reliable fractionator design and show its applicability and potential using fibroblast cells exposed to 50-nm gold nanoparticles. Results & Conclusion: We demonstrate that this approach is suitable for any electron-dense nanomaterial resolvable by electron microscopy and any convex-shaped cells. In addition, the fractionator concept is flexible enough to be used for spatio-temporal or in vivo studies.

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