Real-time, High-throughput Assessment of Nanoparticle Interactions with Single-cells Using Microfluidics

Download or read book Real-time, High-throughput Assessment of Nanoparticle Interactions with Single-cells Using Microfluidics written by Carlota F. A. de Albuquerque Rodrigues da Cunha-Matos and published by . This book was released on 2017 with total page 352 pages. Available in PDF, EPUB and Kindle. Book excerpt: Nanomaterials are increasingly being developed for applications in biotechnology, including the delivery of therapeutic drugs and vaccine antigens. However, there is a lack of screening systems that can rapidly assess nanoparticle uptake and their consequential effects on cells. Established analytical in vitro approaches are often carried out on single time points, rely on time-consuming bulk measurements and are based primarily on populations of immortalised 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 dynamic nature of nanoparticle-cell interactions and/or nanoparticle-mediated compound delivery. The present work addresses these issues by combining microfluidics, nanotechnology and imaging techniques into a high-throughput microfluidic platform to monitor nanoparticle uptake and intracellular processing in real-time and at the single-cell level. For this, a microfluidic device and protocols for cell trapping and live-cell monitoring were developed. In parallel, specific formulations of gold nanorods were produced, tested and optimised for intracellular multimodal imaging. Subsequently, controlled nanorod delivery to cells trapped in the microfluidic array was achieved across a range of concentrations, with intracellular nanorod signal detected using both fluorescence microscopy and surface-enhanced Raman scattering spectroscopy. Furthermore, on-chip measurement of specific cellular responses to nanorod delivery was also demonstrated. As a proof-of-concept application, the potential of the developed platform for understanding nanovaccine delivery and processing was investigated. Controlled delivery of ovalbumin-conjugated gold nanorods to primary dendritic cells was demonstrated, followed by real-time monitoring of nanoparticle uptake and antigen processing 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.