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Ultrasmall, Bright, and Photostable Fluorescent Core–Shell Aluminosilicate Nanoparticles for Live‐Cell Optical Super‐Resolution Microscopy
Advanced Materials ( IF 27.4 ) Pub Date : 2021-01-20 , DOI: 10.1002/adma.202006829 Jacob A Erstling 1, 2 , Joshua A Hinckley 1, 3 , Nirmalya Bag 3 , Jessica Hersh 1 , Grant B Feuer 2 , Rachel Lee 1 , Henry F Malarkey 4 , Fei Yu 1, 3 , Kai Ma 1 , Barbara A Baird 3 , Ulrich B Wiesner 1
Advanced Materials ( IF 27.4 ) Pub Date : 2021-01-20 , DOI: 10.1002/adma.202006829 Jacob A Erstling 1, 2 , Joshua A Hinckley 1, 3 , Nirmalya Bag 3 , Jessica Hersh 1 , Grant B Feuer 2 , Rachel Lee 1 , Henry F Malarkey 4 , Fei Yu 1, 3 , Kai Ma 1 , Barbara A Baird 3 , Ulrich B Wiesner 1
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Stochastic optical reconstruction microscopy (STORM) is an optical super‐resolution microscopy (SRM) technique that traditionally requires toxic and non‐physiological imaging buffers and setups that are not conducive to live‐cell studies. It is observed that ultrasmall (<10 nm) fluorescent core–shell aluminosilicate nanoparticles (aC’ dots) covalently encapsulating organic fluorophores enable STORM with a single excitation source and in a regular (non‐toxic) imaging buffer. It is shown that fourfold coordinated aluminum is responsible for dye blinking, likely via photoinduced redox processes. It is demonstrated that this phenomenon is observed across different dye families leading to probes brighter and more photostable than the parent free dyes. Functionalization of aC’ dots with antibodies allows targeted fixed cell STORM imaging. Finally, aC’ dots enable live‐cell STORM imaging providing quantitative measures of the size of intracellular vesicles and the number of particles per vesicle. The results suggest the emergence of a powerful ultrasmall, bright, and photostable optical SRM particle platform with characteristics relevant to clinical translation for the quantitative assessment of cellular structures and processes from live‐cell imaging.
中文翻译：
用于活细胞光学超分辨率显微镜的超小、明亮、光稳定的荧光核壳铝硅酸盐纳米颗粒
随机光学重建显微镜（STORM）是一种光学超分辨率显微镜（SRM）技术，传统上需要有毒和非生理的成像缓冲液和设置，不利于活细胞研究。据观察，超小（<10 nm）荧光核壳铝硅酸盐纳米颗粒（aC'点）共价封装有机荧光团，使得STORM能够在单一激发源和常规（无毒）成像缓冲液中使用。结果表明，四重配位的铝可能通过光诱导的氧化还原过程导致染料闪烁。事实证明，这种现象在不同的染料家族中都观察到，导致探针比不含母体的染料更亮且更耐光。使用抗体对 aC' 点进行功能化可实现靶向固定细胞 STORM 成像。最后，aC' 点能够实现活细胞 STORM 成像，提供细胞内囊泡大小和每个囊泡颗粒数量的定量测量。结果表明，出现了一种强大的超小、明亮且光稳定的光学 SRM 颗粒平台，其具有与临床转化相关的特征，可用于活细胞成像中细胞结构和过程的定量评估。
更新日期：2021-02-22
中文翻译：
用于活细胞光学超分辨率显微镜的超小、明亮、光稳定的荧光核壳铝硅酸盐纳米颗粒
随机光学重建显微镜（STORM）是一种光学超分辨率显微镜（SRM）技术，传统上需要有毒和非生理的成像缓冲液和设置，不利于活细胞研究。据观察，超小（<10 nm）荧光核壳铝硅酸盐纳米颗粒（aC'点）共价封装有机荧光团，使得STORM能够在单一激发源和常规（无毒）成像缓冲液中使用。结果表明，四重配位的铝可能通过光诱导的氧化还原过程导致染料闪烁。事实证明，这种现象在不同的染料家族中都观察到，导致探针比不含母体的染料更亮且更耐光。使用抗体对 aC' 点进行功能化可实现靶向固定细胞 STORM 成像。最后，aC' 点能够实现活细胞 STORM 成像，提供细胞内囊泡大小和每个囊泡颗粒数量的定量测量。结果表明，出现了一种强大的超小、明亮且光稳定的光学 SRM 颗粒平台，其具有与临床转化相关的特征，可用于活细胞成像中细胞结构和过程的定量评估。
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