Massive wavelength-division multiplexing and imaging with laser particles

使用激光粒子进行大规模波分复用和成像

• 批准号: 9349498
• 负责人: Seok-Hyun Andy Yun
• 金额: $ 116.12万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-30 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9349498
• 关键词: Animal Experiments; Biological Assay; Cell Count; Cells; Cytometry; DNA; Disease; Flow Cytometry; Fluorescence; Fluorescent Dyes; Fluorescent Probes; Fostering; Genes; Genetic; Image; In Vitro; Individual; Injectable; Label; Lasers; Methods; Microscopy; Mission; Mitochondria; Molecular; Mus; Neoplasm Metastasis; Optics; Public Health; Quantum Dots; Reading; Research; Resolution; System; Time; Tissues; United States National Institutes of Health; Virus; Width; biomaterial compatibility; high throughput analysis; improved; in vivo; innovation; instrumentation; intravital microscopy; migration; miniaturize; novel; optical spectra; particle; public health relevance; response; submicron; transcriptome sequencing; treatment response; tumor heterogeneity

ABSTRACT Fluorescence is the current standard method of choice for intravital microscopy and cytometry. However, the broad emission spectrum of fluorescent probes⎯dyes, fluorescent probes, and quantum dots⎯limits the number of cells that can be tracked simultaneously without ambiguity. DNA barcodes can label cells but cannot be visualized in vivo, as they require in vitro genetic reading. The optical principle of stimulated emission and cavity resonance used in a laser can generate extremely narrow spectral line-widths over a broad spectral range. This project will miniaturize lasers to the sizes of mitochondria or viruses and develop instrumentations to utilize the laser particles as novel probes for massively parallel imaging and assays. By tracing individual cells over time in mice, the proliferation, migrations and cell-cell and cell-tissue interactions can be studied in vivo. The cells can be further analyzed by flow cytometry and sorted for gene profiling and single-cell RNA sequencing, providing comprehensive information from molecular, cellular, tissue, and systems levels over millions to billions of cells in a single animal experiment. The first specific aim is to create a new paradigm for imaging-compatible cellular labeling using injectable, biocompatible micro- and submicron-cavity lasers. The second aim is to develop Laser Particle Stimulated Emission (LASE) Microscopy for conducting labeled microscopy in vivo at depths of up to 3 mm. The third specific aim is to demonstrate massively multiplexed, high-throughput cell tracking and analysis. The breakthrough capabilities will be used to dissect tumor heterogeneity in progression, metastasis, and response to therapy at unprecedented single-cell resolution.

抽象的 荧光是当前的浸润显微镜和细胞术的标准选择方法。 然而，荧光探针，荧光探针和 量子点⎯限制了可以简单地跟踪而不会含糊不清的细胞数量。 DNA条形码可以标记细胞，但不能在体内可视化，因为它们需要体外遗传 阅读。激光中使用的刺激发射和空腔共振的光学原理可以 在广泛的光谱范围内产生极狭窄的光谱线宽度。这个项目将 将激光器的小型生物量调至线粒体或病毒和开发工具的尺寸 激光颗粒是大规模平行成像和测定的新问题。通过跟踪 随着时间的流逝，各个细胞在小鼠中，增殖，迁移以及细胞 - 细胞和细胞组织 相互作用可以在体内研究。可以通过流式细胞仪进一步分析细胞，并 分类用于基因分析和单细胞RNA测序，提供全面的信息 从分子，细胞，组织和系统水平超过数百万到数十亿个细胞 动物实验。第一个具体目的是创建一个新的范式，以兼容成像 使用可注射的，生物相容性的微型腔激光器进行细胞标记。第二个 目的是开发用于引导标记的激光颗粒刺激发射（Lase）显微镜 显微镜在体内的深度高达3毫米。第三个具体目的是大规模证明 多路复用，高通量细胞跟踪和分析。突破能力将是 用于在进展，转移和对治疗的反应中剖析肿瘤异质性 空前的单细胞分辨率。