Detection of Silver and TiO2 Nanoparticles in Cells by Flow Cytometry.

Methods in Molecular Biology
Robert Martin Zucker, W K Boyes

Abstract

Evaluation of the potential hazard of man-made nanomaterials has been hampered by a limited ability to observe and measure nanoparticles in cells. A FACSCalibur™ flow cytometer and a Stratedigm S-1000 flow cytometer were used to measure changes in light scatter from cells after incubation with either silver nanoparticles (AgNP) or TiO2 nanoparticles. Within the range of between 0.1 μg/mL and 30 μg/mL the nanoparticles caused a proportional increase of the side scatter and decrease of the forward scatter intensity signals. At the lowest concentrations of TiO2 (ranging between 0.1 μg/mL and 0.3 μg/mL), the flow cytometer can detect as few as 5-10 nanoparticles per cell. The influence of nanoparticles on the cell cycle was detected by nonionic detergent lysis of nanoparticle incubated cells that were stained with DAPI or propidium iodide (PI). Viability of nanoparticle treated cells was determined by PI exclusion. Surface plasmonic resonance (SPR) was detected primarily in the far-red fluorescence detection channels after excitation with a 488 nm laser.Our results suggest that the uptake of nanoparticles within cells can be monitored using flow cytometry. This uptake of nanoparticle data was confirmed by viewing the nanoparticles ...Continue Reading

Citations

Jan 21, 2021·International Journal of Molecular Sciences·Paweł KowalczykIda Franiak-Pietryga
Jul 9, 2021·Ecotoxicology and Environmental Safety·Ju Chang-ChienTsung-Chieh Yao

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