Pulsed Saturated Absorption Competition Microscopy on Nonbleaching Nanoparticles,ACS Photonics

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Pulsed Saturated Absorption Competition Microscopy on Nonbleaching Nanoparticles
ACS Photonics ( IF 6.5 ) Pub Date : 2020-06-09 , DOI: 10.1021/acsphotonics.0c00456
Chuankang Li ₁ , Yuhang Li ₁ , Yubing Han ₁ , Zhimin Zhang ₁ , Yuzhu Li ₁ , Wensheng Wang ₁ , Xiang Hao ₁ , Cuifang Kuang _{1,

2,

3} , Xu Liu _{1,

3}

Affiliation

Super-resolution optical microscopy is a useful tool to investigate physical and biological characteristics on the sub-100-nanometer scale. Here, we develop pulsed saturated absorption competition (pSAC) microscopy to break the diffraction limit in imaging of a series of nonbleaching nanoparticles. Based on the confocal scheme, the input laser beam is divided into doughnut-shaped saturated pulse and Gaussian time-modulated pulse. By adjusting the time sequential of the two pulses, the doughnut-shaped beam transiently saturates the excited-state absorption in the periphery of the focal volume, immediately followed by the Gaussian readout beam which mainly excites fluorophores at the focal center. Through the inhibition of the saturation pulse that confines the fluorescence emission to the central subregion, lock-in detection scheme is utilized to select the characteristic fluorescence signals. We numerically and experimentally demonstrate the resolving capacity of pSAC in imaging biological specimens and subdiffraction resolutions as high as λ/13 and λ/12 are reached in nonfluorescence and fluorescence observation, respectively. The experimental results also show that, compared with stimulated emission depletion (STED) microscopy, pSAC is beneficial to achieve background-suppressed imaging when considering lower-power and fluorescent dye selection.

中文翻译：

非漂白纳米粒子的脉冲饱和吸收竞争显微镜

超分辨率光学显微镜是研究低于100纳米规模的物理和生物学特性的有用工具。在这里，我们开发了脉冲饱和吸收竞争（pSAC）显微镜，打破了一系列非漂白纳米粒子成像的衍射极限。根据共焦方案，将输入激光束分为甜甜圈形饱和脉冲和高斯时间调制脉冲。通过调整两个脉冲的时间顺序，甜甜圈形光束会暂时使焦点体积周围的激发态吸收饱和，紧接着是高斯读出光束，该光束主要在焦点中心激发荧光团。通过抑制将荧光发射限制在中央子区域的饱和脉冲，锁定检测方案用于选择特征荧光信号。我们通过数值和实验证明了pSAC在生物标本成像中的分辨能力，在无荧光和荧光观察中，亚衍射分辨率分别达到λ/ 13和λ/ 12。实验结果还表明，与激发发射耗尽（STED）显微镜相比，pSAC在考虑低功率和荧光染料选择时有利于实现背景抑制的成像。

更新日期：2020-07-15

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