pH and Ca²⁺ play important roles in regulating lysosomal activity and lysosome-mediated physiological and pathological processes. However, effective methods for simultaneous determination of pH and Ca²⁺ is the bottleneck. Herein, a single DNA-based FLIM reporter was developed for real-time imaging and simultaneous quantification of pH and Ca²⁺ in lysosomes with high affinity, in which a specific probe for recognition of Ca²⁺ was assembled onto a DNA nanostructure together with pH-responsive and lysosome-targeted molecules. The developed DNA reporter showed excellent biocompatibility and long-term stability up to ∼56 h in lysosomes. Using this powerful tool, it was discovered that pH was closely related to Ca²⁺ concentration in lysosome, whereas autophagy can be regulated by lysosomal pH and Ca²⁺. Furthermore, Aβ-induced neuronal death resulted from autophagy abnormal through lysosomal pH and Ca²⁺ changes. In addition, lysosomal pH and Ca²⁺ were found to regulate the transformation of NSCs, resulting in Rapamycin-induced antiaging.

pH和Ca ²⁺在调节溶酶体活性和溶酶体介导的生理和病理过程中起重要作用。但是，同时测定pH和Ca ^2+的有效方法是瓶颈。在本文中，开发了一种基于DNA的FLIM报告基因，用于实时成像并以高亲和力同时定量溶酶体中的pH和Ca ²⁺，其中将识别Ca ²⁺的特异性探针与pH响应和溶酶体靶向分子。发达的DNA报道分子在溶酶体中显示出优异的生物相容性和长达56小时的长期稳定性。使用此功能强大的工具，发现pH值与Ca ²⁺密切相关溶酶体中的浓度较高，而自噬可以通过溶酶体的pH和Ca ²⁺来调节。此外，Aβ诱导的神经元死亡是由于溶酶体的pH和Ca ²⁺变化引起的自噬异常所致。另外，发现溶酶体的pH和Ca ²⁺调节NSC的转化，从而导致雷帕霉素诱导的抗衰老。