All-optical fluorescence image recovery using modulated Stimulated Emission Depletion.
All-optical fluorescence image recovery using modulated Stimulated Emission Depletion.
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DOI：
10.1039/c0sc00637h
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发表时间：
2011
期刊：
影响因子：
8.4
通讯作者：
Dickson RM
中科院分区：
文献类型：
--
作者：
Fan C;Hsiang JC;Jablonski AE;Dickson RM
迄今为止，选择性恢复所需荧光信号的荧光调制需要仔细的荧光团选择，并结合长寿命光诱导暗态的重复光学恢复。调制受激辐射损耗采用全光方案，通过在任何外部施加的频率下直接光学地减少发射态，消除了对暗态驻留的需要，从而推广了这种调制方案。利用两个重叠的高斯激光光斑与耗尽光束被强度调制，荧光调制很容易实现与耗尽激光的强度控制的耗尽比。选择性图像恢复，否则不可调制的荧光团信号是直接实现通过这种全光调制，和常见的sted退化多光子激发背景很容易被区分。溶液中的珠子和染料以及固定细胞内结合的荧光团都很容易以这种方式成像。
Fluorescence modulation for selective recovery of desired fluorescence signals has to date required careful fluorophore selection combined with repeated optical recovery from long-lived photoinduced dark states. Adapting an all-optical scheme, modulated Stimulated Emission Depletion generalizes such modulation schemes by eliminating the need for dark state residence by directly optically depopulating the emissive state at any externally applied frequency. Using two overlapped Gaussian laser spots with the depletion beam being intensity-modulated, fluorescence modulation is readily achieved with a depletion ratio governed by the intensity of the depleting laser. Selective image recovery of otherwise unmodulatable fluorophore signals is directly achieved through this all-optical modulation, and common STED-degrading multiphoton-excited background is readily discriminated against. Both beads and dyes in solution as well as fluorophores bound within fixed cells are readily imaged in this manner.