State-of-the-art biological nanoparticle tracking instrument for interdisciplinary use

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Most cells release small membrane-bound vesicles (30-100 nm) termed exosomes, a subpopulation of extracellular vesicles (EVs). Exosomes contain nucleic acids and proteins from the intercellular milieu and will interact with target cells to release a payload, which has the potential to stimulate secondary signalling cascades, including gene transcription. Consequently, exosomes have the potential to enact profound biological effects on the target cells, but the impact of this inter-tissue communication on the whole organism is unclear. From a therapeutics perspective, EVs have become increasingly important as biomarkers of diseases, such as cancers. EVs have also garnered interest as a drug delivery tool, as these vesicles are known to specifically target certain tissues. The extraordinary importance of these biologically active vesicles cannot be overstated; however, due to their small size, identifying and quantifying EVs in vivo has proved challenging. The University of Calgary has a thriving exosome research contingent, spanning three departments (Biological Sciences, Cumming School of Medicine, and Veterinary Medicine), and has resulted in many publications. A noted limitation in all these studies was the inability to quantify EV/exosome concentration in biological fluids. This not only curtails the current research capabilities, but also prevents the research from being competitive on the international stage. Nanoparticle tracking analysis (NTA) is a widely used method of quantifying both size and concentration of EVs in a dilute solution, and this technology is the gold standard for exosome research. Additionally, the NTA method can integrate fluorescent detection, allowing the ability to identify subpopulations of exosomes in a single sample. Particle Metrix's Zetaview TWIN uses NTA technology to detect vesicle size and concentration but additionally will detect two different fluorophores. This is a state-of-the-art instrument capable of measuring vesicles from the nanometer to micrometer size, and would be instrumental in producing high-quality, high-throughput data. Overall, this instrument is a necessary addition to support existing infrastructure in place at UC to study exosomal transport. In particular, ZetaView TWIN is necessary to quantify EVs/exosomes from in vivo studies. For example, does a drug treatment change the size, concentration or particular subpopulation of fluorescently tagged exosomes? Will stress or toxicant exposure in wild and domesticated fishes result in changes to the plasma exosome profile? Will the exosome released by microbes influence the gut epithelial cells? These are just a few of the questions that the primary applicant and co-applicants are hoping to answer using the Zetaview TWIN. The acquisition of the equipment will be critical for the next phase of EV research at the University of Calgary, and will support high-quality HQP recruitment and training, as well as facitlitating breakthrough research. **