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18 May 2010 Sedimentation of agglomerated nanoparticles under cell culture conditions studied by image based analysis
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Toxic effects of nanoparticles can be analyzed with alveolar macrophages in vitro. To quantify exposure of cells to particles we analyzed the sedimentation of nanoparticle agglomerates in cell culture medium (MEM) by means of phase contrast microscopy. Particles were suspended by brief ultrasonication in MEM and pipetted into a glass bottom culture dish on the stage of a Nikon-Biostation under cell culture condition. Successive images were captured from the lowermost optical plane and were converted into binary images. The number of agglomerates (N) as well as the particle-covered area (A) were determined by image analyses. Typically, N increased to a maximum value before it partially decayed due to overlapping and/or optical interference of particles, and finally became constant. In contrast, A increased in a monophasic manner. By means of mathematical modeling we identified the endpoint of sedimentation of particle agglomerates, which is an important though a largely neglected event in most cell culture experiments. This endpoint could be calculated from an approximated model function. As the method can be employed in the presence of cells, a parallel evaluation of particle sedimentation and particle uptake appears possible.
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Darius Schippritt, Martin Wiemann, and Hans-Gerd Lipinski "Sedimentation of agglomerated nanoparticles under cell culture conditions studied by image based analysis", Proc. SPIE 7715, Biophotonics: Photonic Solutions for Better Health Care II, 77153A (18 May 2010);

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