Subcellular and in-vivo Nano-Endoscopy.
Subcellular and in-vivo Nano-Endoscopy.
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DOI：
10.1038/srep34400
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发表时间：
2016-10-03
影响因子：
4.6
通讯作者：
Pyayt A
中科院分区：
文献类型：
--
作者：
Cheemalapati SV;Winskas J;Wang H;Konnaiyan K;Zhdanov A;Roth A;Adapa SR;Deonarine A;Noble M;Das T;Gatenby R;Westerheide SD;Jiang RH;Pyayt A
在亚细胞水平上分析单个细胞对于理解疾病和加速药物发现非常重要。纳米内窥镜允许对单个细胞内部进行微创探测。以前已经提出了一些这样的仪器，但它们要么太复杂而无法制造，要么由于信号采集效率低而需要复杂的外部探测器。在这里，我们提出了一种纳米内窥镜，它可以局部激发标记细胞器中的荧光并收集发射信号进行光谱分析。时域有限差分（FDTD）仿真结果表明，优化后的纳米内窥镜锥度轮廓能将光发射和收集定位在~100 nm范围内。这使得信号检测可用于荧光团附近的纳米光子传感。在插入活细胞的单个细胞器后，制造纳米内窥镜并收集所产生的荧光信号。这包括吖啶橙标记的人成纤维细胞细胞核、Hoechst染色的活肝细胞细胞核和MitoTracker Red标记的MDA-MB-231细胞线粒体的信号收集。将内窥镜插入到产生黄色荧光蛋白的秀丽隐杆线虫（Caenorhabditis elegans）中，并收集荧光信号。据我们所知，这是第一次在亚细胞器水平上进行体内局部荧光信号收集的演示。
Analysis of individual cells at the subcellular level is important for understanding diseases and accelerating drug discovery. Nanoscale endoscopes allow minimally invasive probing of individual cell interiors. Several such instruments have been presented previously, but they are either too complex to fabricate or require sophisticated external detectors because of low signal collection efficiency. Here we present a nanoendoscope that can locally excite fluorescence in labelled cell organelles and collect the emitted signal for spectral analysis. Finite Difference Time Domain (FDTD) simulations have shown that with an optimized nanoendoscope taper profile, the light emission and collection was localized within ~100 nm. This allows signal detection to be used for nano-photonic sensing of the proximity of fluorophores. Upon insertion into the individual organelles of living cells, the nanoendoscope was fabricated and resultant fluorescent signals collected. This included the signal collection from the nucleus of Acridine orange labelled human fibroblast cells, the nucleus of Hoechst stained live liver cells and the mitochondria of MitoTracker Red labelled MDA-MB-231 cells. The endoscope was also inserted into a live organism, the yellow fluorescent protein producing nematode Caenorhabditis elegans, and a fluorescent signal was collected. To our knowledge this is the first demonstration of in vivo, local fluorescence signal collection on the sub-organelle level.
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影响因子：
7.4
作者：
Clark, HA;Kopelman, R;Philbert, MA
通讯作者：
Philbert, MA
影响因子：
38.3
作者：
Yan, Ruoxue;Park, Ji-Ho;Yang, Peidong
通讯作者：
Yang, Peidong
影响因子：
15
作者：
Kasili, PM;Song, JM;Vo-Dinh, T
通讯作者：
Vo-Dinh, T
影响因子：
3.8
作者：
Ladanov, Mikhail;Cheemalapati, Surya;Pyayt, Anna
通讯作者：
Pyayt, Anna
影响因子：
2.2
作者：
Puygranier, BAF;Dawson, P
通讯作者：
Dawson, P