使用基于 TBET 和 AIE 的比例双光子粘度探针跟踪自噬过程,Analytical Chemistry

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使用基于 TBET 和 AIE 的比例双光子粘度探针跟踪自噬过程
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-10-19 , DOI: 10.1021/acs.analchem.2c03555
Shengze Su ₁ , Li Chai ₁ , Qian An ₁ , Wei Hu ₂ , Lina Wang ₂ , Xingcan Li ₁ , Huijuan Zhang ₁ , Chunya Li ₁

Affiliation

自噬是一种细胞自降解过程，在细胞健康和功能中起着关键作用。由于自噬障碍与多种疾病相关，因此检测自噬非常重要。本研究旨在建立基于双传感机制的比例粘度敏感溶酶体靶向双光子荧光探针Vis-sun通过键能转移 (TBET) 和聚集诱导发光 (AIE) 相结合来跟踪自噬过程（自噬过程中溶酶体粘度的增加）。TBET的引入不仅克服了背景信号的干扰，而且实现了两个发射峰的基线分离，从而减少了发射之间的串扰，以及生物目标的无创生物传感和长期实时示踪成像通过引入 AIE。_{体外实验表明，随着水与四氢呋喃体积比（}V_水/ VTHF）的增加，485nm处的荧光强度逐渐降低), 而 605 nm 处的荧光强度显着增加。此外，当水含量达到 100% 时，荧光信号最大。同时，探针表现出对环境粘度的显着依赖性。因此，通过用吗啉实施Vis-sun溶酶体锚定，成功实现了自噬过程中溶酶体粘度的动态监测和中风引起的神经炎症过程中自噬波动的原位成像。

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Tracking Autophagy Process with a TBET and AIE-Based Ratiometric Two-Photon Viscosity Probe

Autophagy is a cellular self-degrading process that plays a key role in cellular health and functioning. Since autophagy disorder is related to many diseases, it is highly important to detect autophagy. This study aimed to establish a dual-sensing mechanism-based ratiometric viscosity-sensitive lysosome-targeted two-photon fluorescent probe Vis-sun to track the autophagy process (the increase in lysosome viscosity during autophagy) by combining through bond energy transfer (TBET) and aggregation-induced emission (AIE). The introduction of TBET not only overcame the interference of background signals but also achieved the baseline separation of two emission peaks, thus reducing the crosstalk between emissions, as well as the noninvasive bio-sensing of biological targets and long-term real-time tracer imaging by introducing AIE. In vitro experiments showed that the fluorescence intensity at 485 nm decreased gradually on increasing the volume ratio of water to tetrahydrofuran (V_water/V_THF), while the fluorescence intensity at 605 nm increased significantly. Also, the fluorescence signal was maximized when the water content reached 100%. At the same time, the probe exhibited a significant dependence on the ambient viscosity. Therefore, the dynamic monitoring of lysosome viscosity during autophagy and the in situ imaging of autophagy fluctuations during stroke-induced neuroinflammation were successfully achieved by implementing Vis-sun lysosome anchoring with morpholine.

更新日期：2022-10-19

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