Self-activated photoblinking of nitrogen vacancy centers in nanodiamonds (sandSTORM): A method for rapid single molecule localization microscopy with unlimited observation time
Self-activated photoblinking of nitrogen vacancy centers in nanodiamonds (sandSTORM): A method for rapid single molecule localization microscopy with unlimited observation time
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DOI：
10.1101/2020.05.20.106716
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发表时间：
2020-05
期刊：
影响因子：
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通讯作者：
Kaarjel K. Narayanasamy;J. Price;Raquel Mesquita-Riberio;M. Mather;I. Jayasinghe
中科院分区：
文献类型：
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作者：
Kaarjel K. Narayanasamy;J. Price;Raquel Mesquita-Riberio;M. Mather;I. Jayasinghe
随机光学重建显微镜（STORM）是最常用的超分辨率显微镜技术之一。 STORM 的流行实现利用芳香族荧光团，并包含许多内在限制，例如染料的有限光稳定性、对非生理氧化还原缓冲液的依赖以及最终受到光闪烁“解离”速率限制的速度。基于自激活纳米金刚石的 STORM (sandSTORM) 已被开发为一种加速 STORM 协议，可收获快速、高量子产率和持续的纳米金刚石 (ND) 光闪烁。 NV0 和 NV- 之间氮空位 (NV) 中心的随机电荷态相互转换发出的光致发光是在无限的成像持续时间内使用传统的 STORM 优化硬件和图像处理协议进行局部化的。与传统 STORM 相比，这可以以大约 3 倍的速度和大约 100 倍的样品光照减少产生分辨率匹配的超分辨率图像。 NDs 已被用于通过免疫标记绘制骨骼肌组织中的兰尼碱受体阵列，并通过 NDs 的内吞作用直接可视化活神经元的内部空间。本文详细介绍了 sandSTORM 的物理基础、优化其性能的因素以及使其成为适用于成像纳米级亚细胞结构的强大 STORM 协议的关键特征。
Stochastic optical reconstruction microscopy (STORM) is one of the most commonly used super-resolution microscopy techniques. Popular implementations of STORM utilize aromatic fluorophores and consist of a number of intrinsic limitations such the finite photostability of the dyes, the reliance upon non-physiological redox buffers and speed which is ultimately limited by the ‘off’-rates of the photoblinking. Self-activated nanodiamond-based STORM (sandSTORM) has been developed as an accelerated STORM protocol which harvests the rapid, high quantum-yield and sustained photoblinking of nanodiamonds (ND). Photoluminescence emanating from the stochastic charge-state interconversion of Nitrogen Vacancy (NV) centers between NV0and NV- is localized using conventional STORM-optimized hardware and image processing protocols over an unlimited duration of imaging. This produces super-resolution images of matching resolution at ∼ 3-times the speed and ∼ 100 times less light exposure to the sample compared to traditional STORM. The enabling NDs have been used to map arrays of ryanodine receptor in skeletal muscle tissues via immunolabelling and directly visualize the internal spaces of living neurons via endocytosis of NDs. This paper details the physical basis of sandSTORM, factors which optimize its performance, and key characteristics which make it a powerful STORM protocol suitable for imaging nanoscale sub-cellular structures.