In vivo multispectral, multiparameter, photoacoustic lymph flow cytometry with natural cell focusing, label-free detection and multicolor nanoparticle probes.
In vivo multispectral, multiparameter, photoacoustic lymph flow cytometry with natural cell focusing, label-free detection and multicolor nanoparticle probes.
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DOI：
10.1002/cyto.a.20587
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发表时间：
2008-10
期刊：
影响因子：
3.7
通讯作者：
Zharov, Vladimir P.
中科院分区：
文献类型：
--
作者：
Galanzha, Ekaterina I.;Shashkov, Evgeny V.;Tuchin, Valery V.;Zharov, Vladimir P.
关键词：
与血液检测相比，淋巴系统中的细胞评估尚未完善确立。这项工作的目标是利用流式细胞术原理开发体内淋巴检测方法。
通过激光（420 - 2300纳米）在由淋巴瓣流体动力学聚焦成单列流的单个细胞中产生光声（PA）信号来对活体动物中的细胞进行计数，并利用内源性吸收作为细胞特异性内在标记，或使用金纳米棒、纳米壳和碳纳米管作为多色探针。光声数据通过高速透射、光热和荧光成像进行了验证。
体内淋巴系统中正常、凋亡和坏死阶段的黑色素瘤细胞及免疫相关细胞计数被证明具有前所未有的灵敏度，可在数百万个正常细胞中检测到一个转移细胞。利用多色标记以及不同波长和时间延迟的激光脉冲实现了对流动细胞的时间分辨光声光谱识别。
多光谱、非侵入性、便携式流式细胞仪可用于动物的临床前研究，并有可能应用于人类，用于无色淋巴管和前哨淋巴结的体内光声成像，同时进行单细胞检测和转移评估，无需标记或使用造影染料和/或具有不同吸收光谱的新型低毒多色探针。
Compared to blood tests, cell assessment in lymphatics is not well-established. The goal of this work was to develop in vivo lymph tests using the principles of flow cytometry. Cells in living animals were counted by laser (420-2300 nm) generation of photoacoustic (PA) signals in individual cells hydrodynamically focused by lymph valves into a single file flow, and using endogenous absorption as intrinsic cell-specific markers, or gold nanorods, nanoshells, and carbon nanotubes as multicolor probes. PA data was verified by high-speed transmission, photothermal, and fluorescent imaging. Counting melanoma and immune-related cells in normal, apoptotic, and necrotic stages in lymphatics in vivo was demonstrated to have the unprecedented sensitivity as one metastatic cell among millions of normal cells. The time-resolved PA spectral identification of flowing cells was achieved using multicolor labels and laser pulses of different wavelengths and time delays. Multispectral, non-invasive, portable flow cytometery can be used for preclinical studies on animals with the potential of translation to humans for in vivo PA mapping of colorless lymph vessels and sentinel nodes with simultaneous single cell detection and metastasis assessment without labeling or use of contrast dyes and/or novel low-toxic multicolor probes with different absorption spectra.
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影响因子：
82.9
作者：
Contag, PR;Olomu, IN;Contag, CH
通讯作者：
Contag, CH
影响因子：
2.2
作者：
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3.5
作者：
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作者：
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通讯作者：
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