Stimulated emission depletion (STED) nanoscopy can achieve super‐resolution imaging of microstructures at the nanoscale by confining the fluorescence emission of a diffraction‐limited focusing spot. The depletion wavelength is typically at the red tail of the emission spectrum of the fluorophore to avoid re‐excitation; however, inefficient fluorescence inhibition on account of the small stimulated emission cross‐section results in a huge demand for the depletion of laser power. Here, an accurate photon extraction method is proposed to eliminate re‐excitation caused by blue shift of the depletion wavelength. After only extracting the photons unaffected by a donut‐shaped depletion laser in the phasor plot, both the resolution and signal‐to‐noise ratio of the STED images are significantly improved. As a result, the increased depletion efficiency at a blue shifted depletion wavelength reduces the power demand for achieving a certain resolution. This approach, with flexible wavelength selectivity owing to the independence of depletion power, made more fluorophores applicable for STED super‐resolution imaging, thereby offering more convenient and efficient service to researchers.

中文翻译：

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在相量图中基于光子提取的激励排放耗尽纳米技术中的再激励消除

通过限制衍射极限聚焦点的荧光发射，受激发射损耗（STED）纳米显微镜可以实现纳米结构的超分辨率成像。耗尽波长通常在荧光团发射光谱的红色尾部，以避免重新激发。然而，由于受激发射截面小，荧光抑制效率低，导致对激光功率的消耗有巨大的需求。在此，提出了一种精确的光子提取方法，以消除由耗尽波长的蓝移引起的再激发。在相量图中仅提取不受环形炸弹耗尽激光影响的光子后，STED图像的分辨率和信噪比都得到了显着改善。结果是，在蓝移的耗尽波长处增加的耗尽效率降低了实现特定分辨率所需的功率。由于耗尽功率的独立性，这种方法具有灵活的波长选择性，这使得更多的荧光团可用于STED超分辨率成像，从而为研究人员提供了更加便捷，高效的服务。