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基于二氰基异佛尔酮的荧光探针实时高保真追踪溶酶体动力学。

Real-Time and High-Fidelity Tracking of Lysosomal Dynamics with a Dicyanoisophorone-Based Fluorescent Probe.

机构信息

Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, College of Chemistry, Central China Normal University, 152 Luoyu Road, Wuhan 430079, P. R. China.

出版信息

Anal Chem. 2021 Dec 21;93(50):16956-16964. doi: 10.1021/acs.analchem.1c04341. Epub 2021 Dec 7.

Abstract

The development of high-performance probes that can visualize and track the dynamic changes of lysosomes is very important for the in-depth study of lysosomes. Herein, we report that a dicyanoisophorone-based probe (named ) can be used for high-fidelity imaging of lysosomes and lysosomal dynamics. can be easily prepared and shows strong far-red to near-infrared emissions centered at 653 nm in water with a huge Stokes shift (224 nm), high quantum yield (Φ = 0.15), high pa value (∼8.79), and good biocompatibility. also shows good cell permeability and can label lysosomes rapidly with bright fluorescence without a time-consuming washing process before imaging. also possesses good photostability and negligible background, making it effective for long-term and high spatiotemporal resolution (0.44 s of exposure) imaging of lysosomes. Moreover, achieved high-fidelity tracking of lysosomal dynamics at an extremely low concentration (1 nM). Finally, we also demonstrated that could real-time track the interactions of lysosomes with other organelles (damaged mitochondria as a model) and image the drug-escape processes from lysosomes. All of the results show that holds broad prospects in lysosome-related research.

摘要

开发能够可视化和跟踪溶酶体动态变化的高性能探针对于深入研究溶酶体非常重要。在此,我们报告了一种基于二氰异佛尔酮的探针(命名为 ),可用于高保真成像溶酶体和溶酶体动力学。 可以很容易地制备,在水中显示出强的远红到近红外发射,中心在 653nm,具有巨大的斯托克斯位移(224nm)、高量子产率(Φ=0.15)、高 pa 值(∼8.79)和良好的生物相容性。 还表现出良好的细胞通透性,能够快速标记溶酶体,无需在成像前进行耗时的洗涤过程。 还具有良好的光稳定性和可忽略的背景,使其能够有效地进行长时间和高时空分辨率(曝光 0.44 秒)的溶酶体成像。此外, 以极低的浓度(1 nM)实现了对溶酶体动力学的高保真跟踪。最后,我们还证明了 可以实时跟踪溶酶体与其他细胞器(以受损线粒体为模型)的相互作用,并对药物从溶酶体逃逸的过程进行成像。所有结果表明, 在溶酶体相关研究中具有广阔的前景。

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