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

基本信息

批准号：
9767768
负责人：
Seok-Hyun Andy Yun
金额：
$ 116.1万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
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

项目摘要

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.

抽象的荧光是目前活体显微镜和细胞术的标准选择方法。然而，荧光探针的宽发射光谱——染料、荧光探针和量子点——限制了可以同时准确跟踪的细胞数量。 DNA条形码可以标记细胞，但无法在体内可视化，因为它们需要体外遗传激光中使用的受激发射和腔谐振的光学原理。该项目将在宽光谱范围内产生极窄的光谱线宽。将激光器小型化至线粒体或病毒大小，并开发利用仪器激光粒子作为大规模并行成像和分析的新型探针。小鼠体内单个细胞随时间的推移，增殖、迁移以及细胞间和细胞组织可以通过流式细胞术进一步分析细胞的相互作用。分类进行基因分析和单细胞RNA测序，提供全面的信息从分子、细胞、组织和系统水平到单个细胞中的数百万到数十亿个细胞第一个具体目标是创建成像兼容的新范例。使用可注射、生物相容性微腔和亚微米腔激光器进行细胞标记。目标是开发激光粒子受激发射（LASE）显微镜用于进行标记第三个具体目标是大规模展示深度达 3 毫米的活体显微镜。多重、高通量细胞追踪和分析能力将是突破性的。用于剖析肿瘤进展、转移和治疗反应的异质性前所未有的单细胞分辨率。