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Detection of Silver Nanoparticles in Cells by Flow Cytometry Using Light Scattering and Far-red Fluorescence
Citation:
Zucker, R., K. Daniel, E. Massaro, S. Karafas, L. Degn, AND W. Boyes. Detection of Silver Nanoparticles in Cells by Flow Cytometry Using Light Scattering and Far-red Fluorescence. Cytometry Part A. Wiley InterScience, Silver Spring, MD, 83(10):962-972, (2013).
Impact/Purpose:
The rapid development and commercialization of man-made nanomaterials have outpaced information regarding the potential hazards of these materials to the environment, humans, or other organisms (1). This situation has prompted the formation of a National Nanotechnology Initiative, which is focused on promoting the safe and responsible development of these promising new technologies (2), and the development of an associated research strategy for the US Environmental Protection Agency (EPA) Office of Research and Development (3-4). Nanoparticles may pose unique health risks beyond those posed by larger particles of the same material due to their reactivity, small sizes, and increased surface areas (3, 5-6). Research on potential hazards of manufactured nanomaterials presents many technical challenges, one of which is a limited ability to detect and quantify nanoparticles in environmental media, tissues, or cells that may have been exposed to nanomaterials. The development of techniques to identify and characterize nanoparticles in cells and various media would be an asset for evaluating potential risks of manufactured nanomaterials.
Description:
The cellular uptake of different sized silver nanoparticles (l0 nm, 50 nm, and 75nm) coated with polyvinylpyrrolidone (PVP) or citrate in ARPE-19 cells following 24 hour incubation was detected by side scatter through the use of a flow cytometer. A large far red fluorescence signal in the 700-800 nm range was detected in five different flow cytometers after 24 hr incubation with 50 nm and 75 nm particles but not with 10 nm particles. A dose dependent correlation of side scatter and far red fluorescence was observed with both PVP and citrate-coated silver particles. The citrate-coated silver nanoparticles (AgNP) revealed a slightly higher scatter and far red fluorescence than did the PVP coated silver nanoparticles. Morphological evaluation showed the silver particles (50 nm and 75 nm) to be clumped and concentrated around the nucleus.