PuSH - Publication Server of Helmholtz Zentrum München

Tian, F. ; Prina-Mello, A.* ; Estrada, G. ; Beyerle, A. ; Möller, W. ; Schulz, S. ; Kreyling, W.G. ; Stöger, T.

A novel assay for the quantification of internalized nanoparticles in macrophages.

Nanotox. 2, 232-242 (2008)
DOI
Open Access Green as soon as Postprint is submitted to ZB.
One of the most urgent requirements in nanotoxicology is a quantitative assessment of internalized nanomaterials in cells. We present an in vitro assay called 'max-flat' for the measurement of internalized particles in macrophages. Fluorescent polystyrene (PS) beads of diameters 1µm, 500 nm, 200 nm, 100 nm and 20 nm were employded. Different concentrations of fibronectin (FN) coated substrates were tested to achieve a maximal cell spreading area and minimal nucleus height, hence 'max-flat'. We found the cell spreading area depends on FN concentration, and it is independent of particle concentration. An optimal conditon of FN was found at 2.5 x 10-3 mg/ml, and it was selected for the max-flat assay to assess the internalization of PS beads. Cells under these conditions neither generate reactive oxygen species nor show noticeable differences in pro-survival/pro-apoptotic signals. Confocal images were employed for the max-flat assay and we set the interval scanning for a Z-stack as nucleus height divided by particle diameter. The max-flat assay provided a significantly higher number of internalized particles and the saturation is reached faster for nano-scale PS beads. We show how the proposed max-flat assay clearly outperform existing techniques by providing easier, more precise and far more reliable access to the number of internalized nanoparticles in macrophages.
Altmetric
Additional Metrics?
[➜Log in]
Edit extra informations Login
Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Materials science; mechanistic toxicology; quantitative assay and internalized nanomaterials
ISSN (print) / ISBN 1743-5390
e-ISSN 1743-5404
Journal Nanotoxicology
Quellenangaben Volume: 2, Issue: 4, Pages: 232-242 Article Number: , Supplement: ,
Publisher Informa Healthcare
Publishing Place London
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Lung Health and Immunity (LHI)
Institute of Bioinformatics and Systems Biology (IBIS)