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Direct visualization and characterization of nanosized biological particles such as viruses, vesicles and protein aggregates are important for various applications in medicine. Specifically, exosomes (30-150 nm in diameter) gained huge interest due to their potential role as a biomarker in cancer diagnosis and prognosis, however direct detection of these particles is challenging due to their small size. Interferometric microscopy allows detection of these particles without using any labels. We show that visibility of nanoparticles can be enhanced in interferometric microscopy by utilizing defocused images. In this paper, with the proposed method Depth Scanning Correlation, we demonstrate label-free detection of individual exosomes isolated from breast cancer cell culture isolated by using Exosome Total Isolation Chip (ExoTIC). Proposed imaging system combined with an isolation tool, can be used in a wide range of applications, where label-free detection of single biological nanoparticles is needed.

Single particle level visualization of biological nanoparticles such as viruses and exosomes is challenging due to their small size and low dielectric contrast. Fluorescence based methods are highly preferred, however they require labelling which may perturb the functionality of the particle of interest. On the other hand, wide-field interferometric microscopy can be used to detect sub-diffraction limited nanoparticles without using any labels. Here we demonstrate that utilization of defocused images enhances the visibility of nanoparticles in interferometric microscopy and thus improves the detectable size limit. With the proposed method termed as Depth Scanning Correlation (DSC) Interferometric Microscopy, we experimentally demonstrate the detection of sub-35nm dielectric particles without using any labels. Furthermore, we demonstrate direct detection of single exosomes. This label-free and high throughput nanoparticle detection technique can be used to sense and characterize biological particles over a range between a few tens to a few hundred nanometers, where conventional methods are insufficient., Scientific and Technological Research Council of Turkey (TÜBİTAK)

An amendment to this paper has been published and can be accessed via a link at the top of the paper.