Ultra-Bright Fluorescent Probes for Flow Cytometry

用于流式细胞术的超亮荧光探针

• 批准号: 9753286
• 负责人: Jiangbo Yu
• 金额: $ 49.49万
• 依托单位: LAMPROGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-20 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9753286
• 关键词: Acquired Immunodeficiency Syndrome; Antigens; Biological Sciences; Biology; Blood; CD4 Positive T Lymphocytes; Cell Cycle; Cell Lineage; Cell surface; Cells; Characteristics; Chemicals; Development; Diagnostic; Dyes; Energy Transfer; Exhibits; Family; Financial compensation; Flow Cytometry; Fluorescence; Fluorescent Dyes; Fluorescent Probes; Immune; Immunotherapy; Label; Lasers; Malignant Neoplasms; Measures; Medicine; Modernization; Monitor; Motivation; Particle Size; Performance; Phase; Photons; Phycoerythrin; Polymers; Protocols documentation; Publications; Quality Control; Quantum Dots; Reagent; Reporting; Series; Signal Pathway; Signal Transduction; Surface; Time; Vertebral column; Viola; Water; Width; Work; absorption; base; cancer biomarkers; design; high throughput technology; instrument; interest; leukemia/lymphoma; nanoparticle; precision medicine; prognostic; quantum; stem; success; treatment response; two-photon; ultraviolet

Project Summary For modern flow cytometry, high-throughput multiplexing is extremely important because of the great need in analyzing a large number of biomolecules on and in a single cell. The multiplexing performance of flow cytometry is hindered by the probe performance including their fluorescence brightness and emission bandwidth. However, there is renewed interest and effort to develop highly multiplexed (e.g. ~20-50 probes) fluorescence based flow cytometry analysis of cells. For this high degree of multiplexing with fluorescence, both probe brightness and narrow emission are critical - specifically, because of how compensation for spectral spillover works, a very bright probe greatly facilitates multiplexing by minimizing spillover spreading, which is a key driver for extremely bright probes in flow cytometry. The ability to detect low- expression antigens is also another important capability provided by very bright probes. Recently, we and others have developed fluorescent nanoparticles based on semiconducting polymers called Pdots. The motivation of adapting fluorescent semiconducting polymers into nanoparticle labels stems from a number of favorable characteristics, such as large absorptivity, high quantum yield, fast emission rates, and excellent photostability. The resulting Pdots exhibit extraordinarily high fluorescence brightness under both one-photon and two-photon excitations, a factor of 102 - 104 higher than conventional fluorescent dyes, and a factor of 10-103 higher than Qdots depending on the particle size. We have successfully designed and prepared a Pdot series with 405nm excitation. Following on this successful development, we propose to design three additional Pdot series for flow cytometry, excitable at 355nm, 488nm, and 561nm, thus matching the laser wavelengths of existing flow cytometers.

项目摘要 对于现代流式细胞仪来说，高通量多路传输是极其重要的，因为 在单个细胞上和单个细胞中分析大量生物分子的巨大需要。这个 流式细胞术的多路传输性能受到探针性能的影响，包括其 荧光亮度和发射带宽。然而，人们对此重新产生了兴趣和努力。 发展基于荧光的高度多重(例如~20-50个探针)的流式细胞术分析 细胞的数量。对于这种与荧光的高度复合，探测器亮度和 窄辐射是至关重要的--具体地说，因为光谱溢出的补偿 工程，一个非常明亮的探头通过最小化溢出扩散极大地促进了多路复用， 这是流式细胞术中极其明亮的探针的关键驱动因素。检测低音的能力- 表达抗原也是非常明亮的探针提供的另一个重要能力。 最近，我们和其他人开发了基于半导体的荧光纳米颗粒 称为Pdots的聚合物。将荧光半导体聚合物应用于 纳米颗粒标签源于许多有利的特性，例如大吸光度， 量子产额高，发射速率快，光稳定性好。由此产生的Pdots展示 在单光子和双光子激发下都具有极高的荧光亮度， 比传统荧光染料高102-104倍，高10-103倍 而不是Q点，这取决于颗粒的大小。 我们成功地设计并制备了405 nm激发的PDot系列。以下是 这次成功的开发，我们建议为FLOW设计另外三个PDOT系列 细胞术，可在355 nm，488 nm和561 nm激发，从而与 现有的流式细胞仪。