Fluorescent nanofoldamers for multiplex flow cytometry

项目总结/摘要 多色流式细胞术（FC）是一种重要的免疫学分析技术， 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的发展，并监测新的有效性， 免疫疗法，通过使用多个生物标志物鉴定关键的细胞亚群， 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、免疫细胞的表征和相关应答 移植和移植物抗宿主病的治疗，以及疫苗和 免疫疗法，以预防和治疗艾滋病毒感染。增加光谱的数量 不同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 在诊断和管理的条件下，用于样本的大小的准确性 诊断有限或细胞数量低，包括来自细针穿刺的细胞， 腹腔镜活检和脑脊液检查光学和激光的改进 推动了最近FC新荧光团的引入。然而，其中大多数拥有 宽且经常重叠的发射轮廓，很像传统的荧光团。重叠 光谱可以通过带通滤波器和数学滤波器的组合使用来分辨。 补偿（重叠减法）;然而，这种补偿增加了实验误差， 降低了灵敏度并限制了多路复用。因此，即使使用原型64色仪器， 领先的研究人员只能实现一个40个参数的面板-进步， 阅读器技术已经超越了荧光团的可用能力。 NIRCNOSciences正在开发两种用于FC的四吡咯荧光团。这些 荧光团是用于多色FC的优秀候选者，这是由于它们的紫外/紫 激发，长斯托克的位移到红色和近红外光谱，非常窄 排放在目前的项目中，NIRCNET寻求完成一项新技术的开发， 被称为FluoroPods，这使我们能够充分利用我们独特的染料组合。这 基于聚合物的系统使得能够使用具有优异光谱特性但 缺乏足够的水溶性和/或在生物系统中严重淬灭。后 完成该项目后，NIRLENTERS科学公司将创造出一个异常明亮的调色板， 仅使用355和405的FluoroPods将构成22种以上新颜色平台的基础 nm激光器。

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.