School of Chemistry and Chemical Engineering, Shandong University of Technology, Zibo, 255049, PR China.
School of Chemistry and Chemical Engineering, Shandong University of Technology, Zibo, 255049, PR China; College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Shandong Provincial Key Laboratory of Clean Production of Fine Chemicals, Shandong Normal University, Jinan, 250014, PR China.
Talanta. 2020 Mar 1;209:120549. doi: 10.1016/j.talanta.2019.120549. Epub 2019 Nov 13.
We herein develop a novel two-photon fluorescent probe termed L-pH for visualization of lysosomal pH within live cells. L-pH is composed of three moieties, including naphthalimide fluorophore as a fluorescence off-on response moiety, piperazine and morpholine groups as lysosomal targeting and pH responsive sites, as well as a reactive benzyl chloride segment for further lysosomal anchoring. The experimental results demonstrate that L-pH can instantaneously respond to various pH values with high sensitivity and selectivity, and has low cytotoxicity and excellent photostability. The one-photon and two-photon fluorescence imaging data indicate L-pH can preferably accumulate into lysosome and monitor the rise of lysosomal pH changes during myriad cell stress conditions, including heat shock, cell apoptosis and mitophagy. Moreover, L-pH was applied for imaging of pH difference in abdominal tissues of mice. L-pH will be a potential tool for monitoring lysosomal pH variation during lysosome-associated physiological and pathological states.
我们在此开发了一种新型的双光子荧光探针，称为 L-pH，用于可视化活细胞内的溶酶体 pH 值。L-pH 由三个部分组成，包括萘酰亚胺荧光团作为荧光关闭-开启响应部分，哌嗪和吗啉基团作为溶酶体靶向和 pH 响应部位，以及反应性苄基氯部分用于进一步的溶酶体锚定。实验结果表明，L-pH 可以对各种 pH 值进行瞬时、高灵敏度和选择性的响应，并且具有低细胞毒性和优异的光稳定性。单光子和双光子荧光成像数据表明，L-pH 可以优先积累到溶酶体中，并在多种细胞应激条件下监测溶酶体 pH 值变化，包括热休克、细胞凋亡和线粒体自噬。此外，L-pH 还用于小鼠腹部组织 pH 差异的成像。L-pH 将成为监测溶酶体相关生理和病理状态下溶酶体 pH 变化的潜在工具。
