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Single cell level quantification of nanoparticlecell interactions using mass cytometry Angela Ivask, Andrew J Mitchell, Christopher M Hope, Simon C Barry, Enzo Lombi, and Nicolas H. Voelcker Anal. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.analchem.7b01006 • Publication Date (Web): 10 Jul 2017 Downloaded from http://pubs.acs.org on July 10, 2017
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Analytical Chemistry
Single cell level quantification of nanoparticle-cell interactions using mass cytometry Angela Ivask[a],[b], Andrew J. Mitchell[c], Christopher M. Hope[d], Simon C. Barry[d], Enzo Lombi*[a] and Nicolas H. Voelcker*[e-g] [a]
Future Industries Institute, University of South Australia, Mawson Lakes, Australia Laboratory of Environmental Toxicology, National Institute of Chemical Physics and Biophysics, Tallinn, Estonia [c] Materials Characterisation and Fabrication Platform, Melbourne School of Engineering, University of Melbourne, Melbourne, Australia [d] University Department of Paediatrics, University of Adelaide, Women's and Children's Hospital, Adelaide, Australia [e] Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia [f] Melbourne Centre for Nanofabrication, Victorian Node of the Australian National Fabrication Facility, Clayton, Victoria, Australia [g] Commonwealth Scientific and Industrial Research Organisation (CSIRO), Clayton, Victoria, Australia [b]
*
Correspondence to:
[email protected];
[email protected] ABSTRACT: Quantification of cell-associated nanoparticles (NPs) is a paramount question in both nanomedicine and nanotoxicology. Inductively coupled plasma mass spectrometry is a well-established method to resolve cell-associated (metal) NPs in bulk cell populations, however such analysis at single cell level remains a challenge. Here we used mass cytometry, a technique that combines single cell analysis and time-of-flight mass spectrometry, to quantitatively analyze extra- and intracellular silver (Ag) in individual Ag NP exposed human T-lymphocytes. The results revealed significant population heterogeneity: e.g., in lymphocytes exposed to 3 µg of 30 nm branched polyethylene imine coated Ag NPs/mL the extracellularly bound Ag varied from 79 to 560 fg and cellular uptake from 17 to 121 fg. Similar amplitude of heterogeneity was observed in cells exposed to various doses of Ag NPs with other sizes and surface coatings, demonstrating the importance of single cell analysis when studying NP-cell interactions. Although mass cytometry has some shortcomings such as inability to analyze potential transformation or dissolution of NPs in cells, we consider this method as the most promising for quantitative assessment of cell-NP interaction at single cell level. Keywords: silver, nanoparticles, cellular uptake, cell surface binding, CyTOF
One of the key questions in biomedical applications of nanosize particles as well as in nanotoxicology is how to quantitatively analyze cell-associated nanoparticles (NPs).1 To date, a variety of quantitative, semi-quantitative and even qualitative methods including inductively-coupled plasma mass spectrometry (ICP-MS), fluorimetry and fluorescence microscopy, fluorescence activated flow cytometry, electron microscopy and synchrotron x-ray fluorescence microscopy or transmission x-ray microscopy1-4 have been employed to assess the cellular association with NPs or their transformation products. For metallic NPs, the most widely used method is ICP-MS which however is typically performed in “bulk” mode by analysing the total metal concentration in a digested cell population5 and therefore, does not count for intra-population variability that may result from target cell phenotypic heterogeneity. Therefore, preferred method over bulk population measurement is single cell level analysis. Quantitative analysis of (metal) NPs at cellular level can be performed using e.g., single-cell (or, time-resolved) ICP-MS, which has been already used to perform multi-elemental analysis of single bac-
terial and yeast cells.6 However, currently issues such as very low (
