Fluorescent nanofoldamers for multiplex flow cytometry

项目摘要/摘要 多色流式细胞术(FC)是免疫学中一种重要的分析技术 研究和临床医学。Fc是了解疾病不可或缺的工具 在细胞和亚细胞水平上的发展以及监测新的 通过使用多个生物标记物识别关键细胞亚群的免疫疗法 面板。它还在不断增长的临床应用中发挥着关键作用，包括开发 传染病疫苗、免疫细胞的特性及相关反应 移植物抗宿主病的移植和治疗，以及疫苗和 预防和治疗艾滋病毒感染的免疫疗法。增加频谱的数量 独特的荧光团将为研究人员提供更大的灵活性，并为临床医生提供更高水平的 在诊断和处理用于以下情况的样品大小的情况下的准确性 诊断有限或细胞数量较少，包括来自细针抽吸的细胞， 腹腔镜术、核心活检和脑脊液检查。光学和激光技术的进步 推动了最近为FC引入新的荧光团。然而，其中的大多数都拥有 广泛且往往重叠的排放轮廓，很像传统的荧光团。重叠 光谱可以通过组合使用带通滤波器和数学计算来解析 补偿(重叠相减)；然而，这种补偿增加了实验误差， 降低敏感度，限制多路传输。因此，即使有了彩色仪的原型， 主要调查人员只能实施40个参数的小组-进展 阅读器技术已经超过了荧光团的现有能力。 涅槃科学公司正在开发用于FC的两个系列的四吡咯荧光团。这些 由于其紫外光/紫外光，荧光团是用于多色FC的极佳候选者 激发，长斯托克的移动到红色和近红外光谱，并且非常窄 排放。在目前的项目中，涅槃寻求完成一项新技术的开发， 这款产品被称为FluoroPods，使我们能够充分利用我们独特的染料产品组合。这 基于聚合物的系统能够使用具有良好光谱特性的染料，但 缺乏足够的水溶性和/或在生物系统中严重猝灭。vt.在.的基础上 项目完成后，涅槃科学将创造一个格外明亮的调色板 将构成仅使用355和405的22种以上新颜色平台的基础 纳米激光。

PROJECT SUMMARY / ABSTRACT Polychromatic flow cytometry (FC) is a vital analytical technique used in immunology, basic research, and clinical medicine. FC is an indispensable tool for understanding disease development at the cellular and subcellular levels and for monitoring the effectiveness of new immunotherapies by identifying key cellular subpopulations using multiple biomarkers within a panel. It also has a critical role in growing clinical applications including development of vaccines for infectious diseases, characterization of immunological cells and responses related to transplantation and treatment of graft-versus-host disease, and development of vaccines and immune therapies to prevent and treat HIV infections. Increasing the number of spectrally distinct fluorophores will give researchers greater flexibility and give clinicians a higher level of accuracy in the diagnosis and management of conditions where the size of the sample used for diagnosis is limited or cell number is low, including those derived from fine needle aspiration, laparoscopy, core biopsy, and cerebrospinal fluid. Improvements in optics and lasers have driven the recent introduction of new fluorophores for FC. However, most of these possess broad and often overlapping emission profiles, much like traditional fluorophores. Overlapping spectra can be resolved by the combined use of bandpass filters and mathematical compensation (overlap subtraction); however, such compensation increases experiment error, reduces sensitivity, and limits multiplexing. Thus, even with a prototype 64-color instrument, leading investigators have only been able to implement a 40-parameter panel – advances in reader technology have outstripped available capabilities in fluorophores. NIRvana Sciences is developing two families of tetrapyrrole fluorophores for use in FC. These fluorophores are excellent candidates for use in polychromatic FC due to their ultra-violet/violet excitation, long Stoke’s shifts into the red and near-infrared spectrums, and very narrow emissions. In the current project, NIRvana seeks to complete development of a new technology, referred to as FluoroPods, that allows us to fully utilize our unique portfolio of dyes. This polymer-based system enables the use of dyes that possess excellent spectral properties but lack sufficient water solubility and/or are heavily quenched in biological systems. Upon completion of the project, NIRvana Sciences will have created a palette of exceptionally bright FluoroPods that will form the basis of a platform of 22+ new colors using only the 355 and 405 nm lasers.