Ultrafast, temporally stochastic STED nanoscopy of millisecond dynamics
Ultrafast, temporally stochastic STED nanoscopy of millisecond dynamics
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DOI：
10.1038/nmeth.3481
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发表时间：
2015-09-01
期刊：
影响因子：
48
通讯作者：
Hell, Stefan W.
中科院分区：
文献类型：
--
作者：
Schneider, Jale;Zahn, Jasmin;Hell, Stefan W.
具有荧光分子状态的寿命的量级的像素停留时间的电光扫描（> 1，000帧/秒）使得受激发射耗尽（STED）纳米显微术在时间上随机。从一个分子的光子检测随机发生在几个扫描帧之一，和光子的空间起源是已知的亚衍射精度。图像是通过对连续帧进行分箱来构建的，使得时间分辨率可以自由调节。我们展示了纳米尺度的囊泡运动在活果蝇幼虫和细胞摄取的病毒颗粒与5至10毫秒的时间分辨率。
Electro-optical scanning (>1,000 frames/s) with pixel dwell times on the order of the lifetime of the fluorescent molecular state renders stimulated emission depletion (STED) nanoscopy temporally stochastic. Photon detection from a molecule occurs stochastically in one of several scanning frames, and the spatial origin of the photon is known with subdiffraction precision. Images are built up by binning consecutive frames, making the time resolution freely adjustable. We demonstrated nanoscopy of vesicle motions in living Drosophila larvae and the cellular uptake of viral particles with 5- to 10-ms temporal resolution.