Label-Free Identification of Persistent Particles in Association with Primary Immune Cells by Imaging Flow Cytometry

The frequency of human exposure to persistent particles via consumer products, air pollution, and work environments is a modern-day hazard and an active area of research. Particle density and crystallinity, which often dictate their persistence in biological systems, are associated with strong light absorption and reflectance. These attributes allow several persistent particle types to be identified without the use of additional labels using laser light-based techniques such as microscopy, flow cytometry, and imaging flow cytometry. This form of identification allows the direct analysis of environmental persistent particles in association with biological samples after in vivo studies and real-life exposures. Microscopy and imaging flow cytometry have progressed with computing capabilities and fully quantitative imaging techniques can now plausibly detail the interactions and effects of micron and nano-sized particles with primary cells and tissues. This chapter summarises studies which have utilized the strong light absorption and reflectance characteristics of particles for their detection in biological specimens. This is followed by the description of methods for the analysis of whole blood samples and the use of imaging flow cytometry to identify particles in association with primary peripheral blood phagocytic cells, using brightfield and darkfield parameters.