Quantitative detection of gold nanoparticles on individual, unstained cancer cells by scanning electron microscopy

Hartsuiker, Liesbeth; Es, Peter van; Petersen, Wilma; Leeuwen, Ton G. van; Terstappen, Leon W.M.M.; Otto, Cees

doi:10.1111/j.1365-2818.2011.03528.x

Cited by 10 publications

(7 citation statements)

References 27 publications

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“…Previous studies have recognized that CeO 2 is biocompatible, and even biofunctional with antioxidant and anti-inflammatory properties. [28][29][30][31][32] Cells dosed with titanium oxide maintained cell viability of 93 AE 1% with 100 ppm, and with 250 ppm cell viability was recovered to 89.8 AE 1.5%, in comparison with 77 AE 0.7% at 3 h of dosing (Fig. 6A).…”

Section: Characterization Of Metal Oxide Nanoparticlesmentioning

confidence: 92%

Imaging interactions of metal oxide nanoparticles with macrophage cells by ultra-high resolution scanning electron microscopy techniques

et al. 2012

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Use of engineered metal oxide nanoparticles in a plethora of biological applications and custom products has warned about some possible dose-dependent cytotoxic effects. Macrophages are key components of the innate immune system used to study possible toxic effects and internalization of different nanoparticulate materials. In this work, ultra-high resolution field emission scanning electron microscopy (FE-SEM) was used to offer new insights into the dynamical processes of interaction of nanomaterials with macrophage cells dosed with different concentrations of metal oxide nanoparticles (CeO2, TiO2 and ZnO). The versatility of FE-SEM has allowed obtaining a detailed characterization of processes of adsorption and endocytosis of nanoparticles, by using advanced analytical and imaging techniques on complete unstained uncoated cells, including secondary electron imaging, high-sensitive backscattered electron imaging, X-ray microanalysis and stereoimaging. Low voltage BF/DF-STEM confirmed nanoparticle adsorption and internalization into endosomes of CeO2 and TiO2, whereas ZnO develop apoptosis after 24 h of interaction caused by dissolution and invasion of cell nucleus. Ultra-high resolution scanning electron microscopy techniques provided new insights into interactions of inorganic nanoparticles with macrophage cells with high spatial resolution.

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Section: Characterization Of Metal Oxide Nanoparticlesmentioning

confidence: 92%

Imaging interactions of metal oxide nanoparticles with macrophage cells by ultra-high resolution scanning electron microscopy techniques

et al. 2012

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“…An alternative approach to eliminate charging effects was applied in the study of gold nanoparticles adsorption on unstained and uncoated cancer cells (Hartsuiker et al, 2011). Cells were grown on a glass precoated with a thin gold layer.…”

Section: Introductionmentioning

confidence: 99%

High resolution SEM imaging of gold nanoparticles in cells and tissues

Goldstein

Soroka

Frušić-Zlotkin

et al. 2014

Journal of Microscopy

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The growing demand of gold nanoparticles in medical applications increases the need for simple and efficient characterization methods of the interaction between the nanoparticles and biological systems. Due to its nanometre resolution, modern scanning electron microscopy (SEM) offers straightforward visualization of metallic nanoparticles down to a few nanometre size, almost without any special preparation step. However, visualization of biological materials in SEM requires complicated preparation procedure, which is typically finished by metal coating needed to decrease charging artefacts and quick radiation damage of biomaterials in the course of SEM imaging. The finest conductive metal coating available is usually composed of a few nanometre size clusters, which are almost identical to the metal nanoparticles employed in medical applications. Therefore, SEM monitoring of metal nanoparticles within cells and tissues is incompatible with the conventional preparation methods. In this work, we show that charging artefacts related to non-conductive biological specimen can be successfully eliminated by placing the uncoated biological sample on a conductive substrate. By growing the cells on glass pre-coated with a chromium layer, we were able to observe the uptake of 10 nm gold nanoparticles inside uncoated and unstained macrophages and keratinocytes cells. Imaging in back scattered electrons allowed observation of gold nanoparticles located inside the cells, while imaging in secondary electron gave information on gold nanoparticles located on the surface of the cells. By mounting a skin cross-section on an improved conductive holder, consisting of a silicon substrate coated with copper, we were able to observe penetration of gold nanoparticles of only 5 nm size through the skin barrier in an uncoated skin tissue. The described method offers a convenient modification in preparation procedure for biological samples to be analyzed in SEM. The method provides high conductivity without application of surface coating and requires less time and a reduced use of toxic chemicals.

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“…Individual nanoparticles are not distinguishable with these conventional imaging techniques, because of low contrast and since the limitation of resolution is around 200 nm. 30 New generation Field Emission Scanning Electron Microscopes (FE-SEM) offers ultra-high resolution. With this advanced imaging technique, it is possible to analyze a number of interactions, adsorption and uptake of small metallic nanoparticles by cells.…”

Section: Introductionmentioning

confidence: 99%

Advanced microscopy of star-shaped gold nanoparticles and their adsorption-uptake by macrophages

et al. 2013

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Metallic nanoparticles have diverse applications in biomedicine, as diagnostics, image contrast agents, nanosensors and drug delivery systems. Anisotropic metallic nanoparticles possess potential applications in cell imaging and therapy+diagnostics (theranostics), but controlled synthesis and growth of these anisotropic or branched nanostructures has been challenging and usually require use of high concentrations of surfactants. Star-shaped gold nanoparticles were synthesized in high yield through a seed mediated route using HEPES as a precise shape-directing capping agent. Characterization was performed using advanced electron microscopy techniques including atomic resolution TEM, obtaining a detailed characterization of nanostructure and atomic arrangement. Spectroscopy techniques showed that particles have narrow size distribution, monodispersity and high colloidal stability, with absorbance into NIR region and high efficiency for SERS applications. Gold nanostars showed to be biocompatible and efficiently adsorbed and internalized by macrophages, as revealed by advanced FE-SEM and backscattered electron imaging techniques of complete unstained uncoated cells. Additionally, low voltage STEM and X-ray microanalysis revealed the ultra-structural location and confirmed stability of nanoparticles after endocytosis with high spatial resolution.

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Quantitative detection of gold nanoparticles on individual, unstained cancer cells by scanning electron microscopy

Cited by 10 publications

References 27 publications

Imaging interactions of metal oxide nanoparticles with macrophage cells by ultra-high resolution scanning electron microscopy techniques

Imaging interactions of metal oxide nanoparticles with macrophage cells by ultra-high resolution scanning electron microscopy techniques

High resolution SEM imaging of gold nanoparticles in cells and tissues

Advanced microscopy of star-shaped gold nanoparticles and their adsorption-uptake by macrophages

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