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Published on: March 28, 2014
Detection of Silver and TiO2 Nanoparticles in Cells by Flow Cytometry
Robert Martin Zucker1, William K Boyes2
1Reproductive and Developmental Toxicology Branch, Public Health and Integrated Toxicology Division, Center for Public Health and Environmental Assessment, Office of Research and Development U.S. Environmental Protection Agency, Research Triangle Park, NC, USA. zucker.robert@epa.gov.
Flow cytometry can now measure nanoparticle uptake in cells, aiding hazard evaluation. This method tracks nanoparticle dose and cellular health, improving nanomaterial safety assessments.
Area of Science:
- Nanotechnology
- Cell Biology
- Toxicology
Background:
- Assessing the hazards of engineered nanomaterials is challenging due to difficulties in observing and quantifying nanoparticles within cells.
- Existing methods for nanoparticle detection in biological systems are often limited in scope and efficiency.
Purpose of the Study:
- To evaluate the potential of flow cytometry for monitoring nanoparticle uptake and cellular effects.
- To establish a flow cytometry-based method for assessing nanoparticle dose-response relationships and cellular toxicity.
Main Methods:
- Utilized FACSCalibur™ and Stratedigm S-1000 flow cytometers to measure light scatter changes in cells exposed to silver nanoparticles (AgNP) and TiO2 nanoparticles.
- Assessed nanoparticle influence on the cell cycle using DAPI or propidium iodide (PI) staining after nonionic detergent lysis.
- Determined cell viability through PI exclusion and detected surface plasmonic resonance (SPR) using far-red fluorescence channels.
Main Results:
- Nanoparticles induced a dose-dependent increase in side scatter and decrease in forward scatter signals.
- Flow cytometry detected as few as 5-10 TiO2 nanoparticles per cell at low concentrations (0.1–0.3 μg/mL).
- Flow cytometry successfully monitored nanoparticle uptake, confirmed by dark-field microscopy, and assessed cellular health.
Conclusions:
- Flow cytometry provides a viable method for monitoring nanoparticle uptake within cells.
- This approach facilitates rapid assessment of nanoparticle dose-response and cellular toxicity, addressing a critical need in nanomaterial safety evaluation.
