Dark-field hyperspectral microscopy for label-free microplastics and nanoplastics detection and identification in vivo: A Caenorhabditis elegans study.

Microplastics pollution is a serious ecological threat, severely affecting environments and human health. Tackling microplastics pollution requires an effective methodology to detect minute polymer particles in environmental samples and organisms. Here were report a novel methodology to visualise and identify nanoscale (down to 100 nm) and microscale synthetic commercially-available uniform spherical polymer particles using dark-field hyperspectral microscopy in visible-near infrared (400-1000 nm) wavelength range. Polystyrene particles with diameters between 100 nm-1 μm, polymethacrylate 1 μm and melamine formaldehyde 2 μm microspheres suspended in pure water samples were effectively imaged and chemically identified based on spectral signatures and image-assisted analysis. We succeeded in visualisation and spectral identification of pure and mixed nano- and microplastics in vivo employing optically-transparent Caenorhabditis elegans nematodes as a model to demonstrate the ingestion and tissue distribution of microplastics. As we demonstrate here, dark-field hyperspectral microscopy is capable for differentiating between chemically-different microplastics confined within live invertebrate intestines. Moreover, this optical technology allows for quantitative identification of microplastics ingested by nematodes. We believe that this label-free non-destructive methodology will find numerous applications in environmental nano- and microplastics detection and quantification, investigation of their biodistribution in tissues and organs and nanotoxicology.