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Proceedings Paper

Optical analysis of nanomaterial-cell interactions: flow cytometry and digital holographic microscopy
Author(s): Sarah Mues; Jan Antunovic; Rainer Ossig; Björn Kemper; Jürgen Schnekenburger
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Paper Abstract

The in vitro cytotoxicity assessment of engineered nanoparticles commonly involves the measurement of different endpoints like the formation of reactive oxygen species, cell viability or cell death. Usually these parameters are determined by optical readouts of enzymatically converted substrates that often interfere with the tested nanomaterials. Using cell viability (WST-8) and cell death (LDH) as parameter we have initially investigated the toxic effects of spherical (NM 300) and rod shaped (NM 302) silver nanomaterials with a matrix of four cell lines representing different functions: lung and kidney epithelial cells, macrophages and fibroblasts. In addition, we have used a label-free flow cytometer configuration to investigate interactions of particles and macrophages by side scatter signal analysis. Finally, we explored digital holographic microscopy (DHM) for multimodal label-free analysis of nanomaterial toxicity. Quantitative DHM phase images were analyzed for cell thickness, volume, density, dry mass and refractive index. We could demonstrate that silver spheres lead to more cytotoxic effects than rods in all four examined cell lines and both assay. Exemplarily a dose dependent interaction increase of cells with NM 300 and NM 302 analyzed by flow cytometry is shown. Furthermore, we found that the refractive index of cells is influenced by incubation with NM 300 in a decreasing manner. A 24 hours time-lapse measurement revealed a dose dependent decrease of dry mass and surface area development indicating reduced cell viability and cell death. Our results demonstrate digital holographic microscopy and flow cytometry as valuable label-free tools for nanomaterial toxicity and cell interaction studies.

Paper Details

Date Published: 22 June 2015
PDF: 6 pages
Proc. SPIE 9529, Optical Methods for Inspection, Characterization, and Imaging of Biomaterials II, 95290E (22 June 2015); doi: 10.1117/12.2184869
Show Author Affiliations
Sarah Mues, Westfälische Wilhelms-Univ. Münster (Germany)
Jan Antunovic, Westfälische Wilhelms-Univ. Münster (Germany)
Rainer Ossig, Westfälische Wilhelms-Univ. Münster (Germany)
Björn Kemper, Westfälische Wilhelms-Univ. Münster (Germany)
Jürgen Schnekenburger, Westfälische Wilhelms-Univ. Münster (Germany)

Published in SPIE Proceedings Vol. 9529:
Optical Methods for Inspection, Characterization, and Imaging of Biomaterials II
Pietro Ferraro; Simonetta Grilli; Monika Ritsch-Marte; David Stifter, Editor(s)

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