Massive wavelength-division multiplexing and imaging with laser particles
Massive wavelength-division multiplexing and imaging with laser particles
批准号:
9349498
负责人:
Seok-Hyun Andy Yun
金额:
$116.12万
依托单位国家:
美国
项目类别:
财政年份:
2016
资助国家:
美国
项目状态:
已结题
起止时间:
2016-09-30 至 2021-07-31
关键词:
Animal ExperimentsBiological AssayCell CountCellsCytometryDNADiseaseFlow CytometryFluorescenceFluorescent DyesFluorescent ProbesFosteringGenesGeneticImageIn VitroIndividualInjectableLabelLasersMethodsMicroscopyMissionMitochondriaMolecularMusNeoplasm MetastasisOpticsPublic HealthQuantum DotsReadingResearchResolutionSystemTimeTissuesUnited States National Institutes of HealthVirusWidthbiomaterial compatibilityhigh throughput analysisimprovedin vivoinnovationinstrumentationintravital microscopymigrationminiaturizenoveloptical spectraparticlepublic health relevanceresponsesubmicrontranscriptome sequencingtreatment responsetumor heterogeneity
中文摘要
摘要
荧光是目前活体显微镜和细胞学的标准选择方法。
然而,荧光探针的广泛发射光谱⎯染料、荧光探针和
量子点⎯限制了可以同时跟踪的细胞数量,而不会有歧义。
DNA条形码可以标记细胞,但不能在体内可视化,因为它们需要体外基因
阅读。用于激光器的受激发射和腔内谐振的光学原理
在很宽的光谱范围内产生极窄的谱线宽度。这个项目将
将激光小型化到线粒体或病毒的大小,并开发可利用的仪器
激光粒子作为大规模并行成像和分析的新型探针。通过跟踪
小鼠体内单个细胞随时间的变化、增殖、迁移以及细胞-细胞和细胞组织
相互作用可以在体内进行研究。这些细胞可以通过流式细胞仪进行进一步分析
对基因图谱和单细胞RNA测序进行分类,提供全面的信息
从分子、细胞、组织和系统级别的数百万到数十亿个细胞
动物实验。第一个具体目标是创建与图像兼容的新范例
使用可注入的、生物兼容的微腔和亚微米腔激光器进行细胞标记。第二
目的是发展用于导电标记的激光粒子受激发射(LASE)显微镜
活体显微镜,最深可达3毫米。第三个具体目标是大规模示威
多路复用、高通量小区跟踪和分析。突破性的能力将是
用于分析肿瘤进展、转移和治疗反应的异质性。
史无前例的单细胞分辨率。
英文摘要
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.
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会议论文
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海外基金