title: Real-time assessment of nanoparticle-mediated antigen delivery and cell response.
abstract: Nanomaterials are increasingly being developed for applications in biotechnology, including the delivery of therapeutic drugs and of vaccine antigens. However, there is a lack of screening systems that can rapidly assess the dynamics of nanoparticle uptake and their consequential effects on cells. Established in vitro approaches are often carried out on a single time point, rely on time-consuming bulk measurements and are based primarily on populations of cell lines. As such, these procedures provide averaged results, do not guarantee precise control over the delivery of nanoparticles to cells and cannot easily generate information about the dynamics of nanoparticle-cell interactions and/or nanoparticle-mediated compound delivery. Combining microfluidics and nanotechnology with imaging techniques, we present a microfluidic platform to monitor nanoparticle uptake and intracellular processing in real-time and at the single-cell level. As proof-of-concept application, the potential of such a system for understanding nanovaccine delivery and processing was investigated and we demonstrate controlled delivery of ovalbumin-conjugated gold nanorods to primary dendritic cells. Using time-lapse microscopy, our approach allowed monitoring of uptake and processing of nanoparticles across a range of concentrations over several hours on hundreds of single-cells. This system represents a novel application of single-cell microfluidics for nanomaterial screening, providing a general platform for studying the dynamics of cell-nanomaterial interactions and representing a cost-saving and time-effective screening tool for many nanomaterial formulations and cell types.
authors.affiliation: Department of Biomedical Engineering, University of Strathclyde, 106 Rottenrow, Glasgow, G4 0NW, UK.
authors.forename: Carlota A
authors.initials: CA
authors.lastname: Cunha-Matos
authors.affiliation: Centre for Biophotonics, Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 0RE, UK.
authors.forename: Owain R
authors.initials: OR
authors.lastname: Millington
authors.affiliation: Centre for Molecular Nanometrology, WestCHEM, Department of Pure and Applied Chemistry, Technology and Innovation Centre, University of Strathclyde, 99 George St, Glasgow, G1 1RD, UK.
authors.forename: Alastair W
authors.initials: AW
authors.lastname: Wark
authors.affiliation: Centre for Microsystems and Photonics, Electronic and Electrical Engineering, University of Strathclyde, 204 George Street, Glasgow, G1 1XW, UK. michele.zagnoni@strath.ac.uk.
authors.forename: Michele
authors.initials: M
authors.lastname: Zagnoni
mesh_terms: D065567:Absorption, Physiological; D000818:Animals; D000941:Antigens / Q000008:administration & dosage* / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D016503:Drug Delivery Systems; D004867:Equipment Design; D006046:Gold / Q000737:chemistry*; D057166:High-Throughput Screening Assays / Q000295:instrumentation; D006801:Humans; D007091:Image Processing, Computer-Assisted; D056656:Lab-On-A-Chip Devices; D053768:Metal Nanoparticles / Q000737:chemistry*; D020297:Mice, Congenic; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D043942:Nanotubes / Q000737:chemistry*; D010047:Ovalbumin / Q000008:administration & dosage / Q000378:metabolism; D000075082:Proof of Concept Study; D059010:Single-Cell Analysis; D059008:Time-Lapse Imaging
chemical_list: D000941:Antigens; D006046:Gold; D010047:Ovalbumin