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Main Title: Quantification of silver nanoparticle uptake and distribution within individual human macrophages by FIB/SEM slice and view
Author(s): Gührs, Erik
Schneider, Michael
Günther, Christian M.
Hessing, Piet
Heitz, Karen
Wittke, Doreen
Oliver, Ana López-Serrano
Jakubowski, Norbert
Plendl, Johanna
Eisebitt, Stefan
Haase, Andrea
Type: Article
Language Code: en
Abstract: Background Quantification of nanoparticle (NP) uptake in cells or tissues is very important for safety assessment. Often, electron microscopy based approaches are used for this purpose, which allow imaging at very high resolution. However, precise quantification of NP numbers in cells and tissues remains challenging. The aim of this study was to present a novel approach, that combines precise quantification of NPs in individual cells together with high resolution imaging of their intracellular distribution based on focused ion beam/ scanning electron microscopy (FIB/SEM) slice and view approaches. Results We quantified cellular uptake of 75 nm diameter citrate stabilized silver NPs (Ag 75 Cit) into an individual human macrophage derived from monocytic THP-1 cells using a FIB/SEM slice and view approach. Cells were treated with 10 μg/ml for 24 h. We investigated a single cell and found in total 3138 ± 722 silver NPs inside this cell. Most of the silver NPs were located in large agglomerates, only a few were found in clusters of fewer than five NPs. Furthermore, we cross-checked our results by using inductively coupled plasma mass spectrometry and could confirm the FIB/SEM results. Conclusions Our approach based on FIB/SEM slice and view is currently the only one that allows the quantification of the absolute dose of silver NPs in individual cells and at the same time to assess their intracellular distribution at high resolution. We therefore propose to use FIB/SEM slice and view to systematically analyse the cellular uptake of various NPs as a function of size, concentration and incubation time.
Issue Date: 21-Mar-2017
Date Available: 3-Dec-2018
DDC Class: 610 Medizin und Gesundheit
Subject(s): nanoparticles
FIB/SEM slice and view
absolute dose
cellular internalization
Sponsor/Funder: TU Berlin, Open-Access-Publikationsfonds – 2017
Journal Title: Journal of Nanobiotechnology
Publisher: BioMed Central
Publisher Place: London
Volume: 15
Article Number: 21
Publisher DOI: 10.1186/s12951-017-0255-8
ISSN: 1477-3155
Appears in Collections:FG Röntgenoptik und Nanometer-Optik » Publications

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