No need for beads! Virus-derived biological standards for nanoscale flow cytometry

纳米级流式细胞术(NFC)是一项新兴的革命性技术，旨在分析微小生物颗粒上表达的细胞衍生表面标记的组成，最明显的是病毒和细胞外小泡(EV)。EV类似于由所有类型的细胞不断释放的微小气泡，是协调生长、分化的一种交流手段，也可以警告其他细胞潜在的危险，如损伤、感染和癌症、帕金森氏症和阿尔茨海默氏症等疾病。分析从血液、唾液和尿液等生物液中分离出来的EV的表面标记，可能会提供疾病或感染的早期预警信号。因此，电动汽车表面的标记将成为医疗和兽医临床诊断的有价值和强大的工具。尽管NFC能够分析电动汽车表面标志，但由于缺乏合适的商业仪器校准标准和实验控制，NFC在医院和研究中心的广泛部署和采用一直受到阻碍。我们已经开发出模拟电动汽车大小和物理特性的荧光病毒衍生颗粒，从而成为NFC极其可靠和有效的控制手段。这种病毒可以持续、廉价和大量地生产。这里提出的工作将进一步改进这些病毒，以满足特定的研究和临床要求。预期的结果是生产10种不同的病毒衍生标准或对照，可用于各种研究和临床NFC应用，以准确分析EV表面标志物。我们将通过创建一家致力于制造这些病毒衍生颗粒的初创公司来确保这些颗粒的供应。这家初创公司将向NFC仪器制造商和试剂分销商提供病毒衍生标准，他们将向他们的学术和行业客户提供这些标准。加拿大的经济和社会利益包括培训高素质的人员，以及创建一家初创公司，该公司将生产一种用于NFC应用的试剂，这将反过来导致用于人类、鱼类和牲畜的新一代疾病和感染的非侵入性诊断测试。

Nanoscale flow cytometry (NFC) is an emerging and transformative technology that is designed to analyze the composition of cell-derived surface markers expressed on minute biological particles, most notably viruses and extracellular vesicles (EVs). EVs resemble tiny bubbles constantly released by all cell types and serve as a means of communication to coordinate growth, differentiation, but also to warn other cells of potential dangers such as injury, infection and diseases such as cancer, Parkinson's and Alzheimer's. Analyzing the surface markers of EVs isolated from biological fluids such as blood, saliva and urine may provide early warning signs of disease or infection. As such, markers on the surface of EVs are poised to become valuable and powerful tools for medical and veterinary clinical diagnostics. Despite NFC being capable of analyzing EV surface markers, its broad deployment and adoption by hospitals and research centers has been hindered by the absence of suitable commercial instrument calibration standards and experimental controls. We have developed fluorescent virus-derived particles that mimic the size and physical properties of EVs, and thereby act as extremely reliable and effective controls for NFC. The viruses can be produced consistently, cheaply and in large quantities. The work proposed here will further improve these viruses to meet specific research and clinical requirements. The anticipated outcome is the production of 10 different virus-derived standards or controls that can be used in various research and clinical NFC applications for the accurate analysis of EV surface markers. We will ensure the supply of theses virus-derived particles through the creation of a start-up company dedicated to making them. The start-up will supply NFC instrument manufacturers and reagent distributors with the virus-derived standards, and they will make them available to their academic and industry clients. Economic and societal benefits to Canada include the training of highly qualified personnel, and the creation of a start-up that will manufacture a reagent for NFC applications which will, in turn, lead to a new generation of non-invasive diagnostic tests for disease and infection for use in humans, fish and livestock.**************