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

大多数细胞释放被称为外泌体的小的膜结合囊泡（30-100 nm），外泌体是细胞外囊泡（EV）的亚群。外泌体含有来自细胞间环境的核酸和蛋白质，并将与靶细胞相互作用以释放有效负载，这有可能刺激二级信号级联，包括基因转录。因此，外泌体有可能对靶细胞产生深远的生物学效应，但这种组织间通讯对整个生物体的影响尚不清楚。从治疗学的角度来看，EV作为疾病（如癌症）的生物标志物变得越来越重要。EV作为药物递送工具也引起了人们的兴趣，因为已知这些囊泡特异性靶向某些组织。这些生物活性囊泡的非凡重要性不能被夸大;然而，由于它们的小尺寸，在体内识别和定量EV已被证明具有挑战性。卡尔加里大学拥有一支蓬勃发展的外泌体研究队伍，跨越三个系（生物科学，卡明医学院和兽医学），并发表了许多出版物。所有这些研究中的一个值得注意的限制是无法量化生物流体中的EV/外来体浓度。这不仅限制了目前的研究能力，而且也使研究无法在国际舞台上竞争。纳米颗粒跟踪分析（NTA）是一种广泛使用的定量稀释溶液中EV大小和浓度的方法，该技术是外泌体研究的金标准。此外，NTA方法可以整合荧光检测，从而能够鉴定单个样品中的外来体亚群。Particle Metrix的Zetaview TWIN使用NTA技术来检测囊泡大小和浓度，但还将检测两种不同的荧光团。这是一种最先进的仪器，能够测量从纳米到微米大小的囊泡，并将有助于产生高质量，高通量的数据。总的来说，该仪器是一个必要的补充，以支持现有的基础设施在UC研究外泌体运输。特别是，ZetaView TWIN对于从体内研究中定量EV/外来体是必要的。例如，药物治疗是否会改变荧光标记的外泌体的大小、浓度或特定亚群？野生和驯养鱼类的应激或毒物暴露会导致血浆外泌体谱的变化吗？微生物释放的外泌体会影响肠道上皮细胞吗？这些只是主要申请人和共同申请人希望使用Zetaview TWIN回答的几个问题。这些设备的采购对于卡尔加里大学电动汽车研究的下一阶段至关重要，并将支持高质量的HQP招聘和培训，以及促进突破性研究。**

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. **