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Nanoplastics (NP) are of environmental and human health concern. We tested a novel NP extraction method and scanning transmission X-ray spectro-microscopy (STXM) in combination with near-edge X-ray absorption fine-structure spectroscopy (NEXAFS) to image and identify individual NP in environmental and food matrices. We (1) discussed the potential of STXM compared to other methods potentially suitable for NP analysis, (2) applied the method on NP suspensions of eight of the most common polymers, (3) analyzed environmental water and soil samples spiked with NP and (4) characterized NP in tea water infused in plastic teabags and unspiked soil samples. Here we show that STXM has methodological advantages and that polymers give characteristic spectra, which allows NP identification in environmental and food matrices. For soils we deliver a visual and spectroscopic characterization of NP, proving their presence and highlighting their diversity. Thus, STXM, can be used for the detection and characterisation of NP in different types of matrices.