Live-Cell STED Microscopy with Genetically Encoded Biosensor,Nano Letters

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Live-Cell STED Microscopy with Genetically Encoded Biosensor
Nano Letters ( IF 9.6 ) Pub Date : 2015-04-17 00:00:00 , DOI: 10.1021/nl504710z
Natalia M. Mishina _{1,

2} , Alexander S. Mishin _{1,

2} , Yury Belyaev ₃ , Ekaterina A. Bogdanova ₁ , Sergey Lukyanov _{1,

2} , Carsten Schultz ₃ , Vsevolod V. Belousov _{1,

2}

Affiliation

Of the various super-resolution techniques, stimulated emission depletion (STED) microscopy achieves the best temporal resolution at high spatial resolution, enabling live-cell imaging beyond the diffraction limit. However, STED and most other super-resolution imaging methods utilize a particular type of information extractable from the raw data, namely the positions of fluorophores. To expand on the use of super-resolution techniques, we report here the live-cell STED microscopy of a dynamic biosensor. Using the fluorescent H₂O₂ sensor HyPer2 for subdiffraction imaging, we were able not only to image filaments with superior resolution by localizing emission but also to trace H₂O₂ produced within living cell by monitoring brightness of the probe. STED microscopy of HyPer2 demonstrates potential utility of FP-based biosensors for super-resolution experiments in situ and in vivo.

中文翻译：

带有基因编码生物传感器的活细胞STED显微镜

在各种超分辨率技术中，受激发射损耗（STED）显微镜在高空间分辨率下实现了最佳的时间分辨率，从而使活细胞成像超出了衍射极限。但是，STED和大多数其他超分辨率成像方法利用了可从原始数据中提取的特定类型的信息，即荧光团的位置。为了扩展超分辨率技术的使用，我们在这里报告动态生物传感器的活细胞STED显微镜。使用荧光H ₂ O ₂传感器HyPer2进行亚衍射成像，我们不仅能够通过局部发射来对具有高分辨率的细丝进行成像，而且还能跟踪H ₂ O ₂通过监测探头的亮度在活细胞内产生。HyPer2的STED显微镜证明了基于FP的生物传感器在原位和体内超分辨率实验中的潜在用途。

更新日期：2015-04-17

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