Switchable and Functional Fluorophores for Multidimensional Single-Molecule Localization Microscopy.
Switchable and Functional Fluorophores for Multidimensional Single-Molecule Localization Microscopy.
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DOI：
10.1021/cbmi.3c00045
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发表时间：
2023-08-28
期刊：
影响因子：
--
通讯作者：
Zhang, Yang
中科院分区：
文献类型：
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作者：
Liu, Yunshu;Shahid, Md Abul;Mao, Hongjing;Chen, Jiahui;Waddington, Michael;Song, Ki-Hee;Zhang, Yang
多维单分子定位显微镜（mSMLM）代表了超分辨率显微镜技术领域的范式转变。它通过询问可切换荧光团的发射特性，提供了纳米尺度单分子空间位置和功能信息的同时检测。后者通过精心操纵的激光照明和单分子检测策略精确地调整以报告其局部环境。这个观点强调了mSMLM最近的进展，重点是荧光团设计及其集成到mSMLM成像系统。特别感兴趣的是在同时复用超分辨率成像，纳米级极性和疏水性作图，以及单分子定向成像方面的成就。本文还讨论了mSMLM的挑战和前景，包括开发更具活力和功能的荧光探针，优化光学实现以明智地利用光子预算，以及成像分析和机器学习技术的进步。
Multidimensional single-molecule localization microscopy (mSMLM) represents a paradigm shift in the realm of super-resolution microscopy techniques. It affords the simultaneous detection of single-molecule spatial locations at the nanoscale and functional information by interrogating the emission properties of switchable fluorophores. The latter is finely tuned to report its local environment through carefully manipulated laser illumination and single-molecule detection strategies. This Perspective highlights recent strides in mSMLM with a focus on fluorophore designs and their integration into mSMLM imaging systems. Particular interests are the accomplishments in simultaneous multiplexed super-resolution imaging, nanoscale polarity and hydrophobicity mapping, and single-molecule orientational imaging. Challenges and prospects in mSMLM are also discussed, which include the development of more vibrant and functional fluorescent probes, the optimization of optical implementation to judiciously utilize the photon budget, and the advancement of imaging analysis and machine learning techniques.