Fluorescence-encoded gold nanoparticles: library design and modulation of cellular uptake into dendritic cells.

In order to harness the unique properties of nanoparticles for novel clinical applications and to modulate their uptake into specific immune cells we designed a new library of homo- and hetero-functional fluorescence-encoded gold nanoparticles (Au-NPs) using different poly(vinyl alcohol) and poly(ethylene glycol)-based polymers for particle coating and stabilization. The encoded particles were fully characterized by UV-Vis and fluorescence spectroscopy, zeta potential and dynamic light scattering. The uptake by human monocyte derived dendritic cells in vitro was studied by confocal laser scanning microscopy and quantified by fluorescence-activated cell sorting and inductively coupled plasma atomic emission spectroscopy. We show how the chemical modification of particle surfaces, for instance by attaching fluorescent dyes, can conceal fundamental particle properties and modulate cellular uptake. In order to mask the influence of fluorescent dyes on cellular uptake while still exploiting its fluorescence for detection, we have created hetero-functionalized Au-NPs, which again show typical particle dependent cellular interactions. Our study clearly prove that the thorough characterization of nanoparticles at each modification step in the engineering process is absolutely essential and that it can be necessary to make substantial adjustments of the particles in order to obtain reliable cellular uptake data, which truly reflects particle properties.

Quantification of nanoparticles at the single-cell level: an overview about state-of-the-art techniques and their limitations.

With the increasing production and use of engineered nanoparticles it is crucial that their interaction with biological systems is understood. Due to the small size of nanoparticles, their identification and localization within single cells is extremely challenging. Therefore, various cutting-edge techniques are required to detect and to quantify metals, metal oxides, magnetic, fluorescent, as well as electron-dense nanoparticles. Several techniques will be discussed in detail, such as inductively coupled plasma atomic emission spectroscopy, flow cytometry, laser scanning microscopy combined with digital image restoration, as well as quantitative analysis by means of stereology on transmission electron microscopy images. An overview will be given regarding the advantages of those visualization/quantification systems, including a thorough discussion about limitations and pitfalls.

Evaluation of chocolate as a source of dietary copper

Chocolate has frequently been proposed to be a valuable source of dietary copper, but data on the copper content of major contemporary chocolate brands are scarce. The copper content of 22 brands of chocolate, many of which are sold worldwide, is thus presented here. A reliable hot ashing procedure to determine the copper content of chocolate by inductively coupled plasma atomic emission spectroscopy is also described. It was found that the copper contents of the chocolates analyzed here varied in the range of 1.85 ± 0.10 to 16.50 ± 1.29 μg/g. There was a linear correlation of the copper content of chocolate to its cocoa content with a correlation coefficient R2 of 0.89, showing that the copper was largely contributed to the chocolate by the cocoa. The value of chocolate as a source of dietary copper is discussed.