9 March 2016 Novel optical approaches for label-free quantification of nano-cytotoxic effects
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Abstract
Commonly used cytotoxicity assays to determine the formation of reactive oxygen species, cell viability or cell death are often affected by applied nanomaterials, which lead to false-positive or false-negative results. Thus, novel nanomaterial toxicity testing strategies that allow for high nanomaterial doses to determine Low Effect Levels (LOEL) even of low toxic materials are of high interest. We demonstrate novel approaches to quantify cytotoxic effects with new parameter sets such as cellular refractive index, volume, density and dry mass that are obtained by digital holographic microscopy (DHM). Furthermore, we correlate results obtained from spherical (NM 300) and rod shaped (NM 302) silver nanomaterials with established cell viability and cell death assays. Moreover, in a label-free flow cytometry configuration, cell-nanoparticle-interaction-kinetics were determined by side scatter signal analysis. We demonstrate that silver spheres show a higher cytotoxicity than silver rods and found that this effect correlates with a decrease of the intracellular refractive index and a decreased temporal development of dry mass and cell covered surface area indicating reduced cell viability and increased cell death. Results from side scatter analysis suggest a dose-dependent uptake kinetics of both materials that correlates with cytotoxicity data of the established assays. Taken together, our results demonstrate DHM and flow cytometry as promising novel label-free tools for nanomaterial toxicity and cell particle interaction studies.
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Sarah Mues, Sarah Mues, Jan Antunovic, Jan Antunovic, Steffi Ketelhut, Steffi Ketelhut, Björn Kemper, Björn Kemper, Jürgen Schnekenburger, Jürgen Schnekenburger, "Novel optical approaches for label-free quantification of nano-cytotoxic effects", Proc. SPIE 9719, Biophysics, Biology, and Biophotonics: the Crossroads, 97190J (9 March 2016); doi: 10.1117/12.2213269; https://doi.org/10.1117/12.2213269
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