Neural stem cells (NSCs) must exit quiescence to produce neurons; however, our understanding of this process remains constrained by the technical limitations of current technologies. Fluorescence lifetime imaging (FLIM) of autofluorescent metabolic cofactors has been used in other cell types to study shifts in cell states driven by metabolic remodeling that change the optical properties of these endogenous fluorophores. Using this non-destructive, live-cell, and label-free strategy, we found that quiescent NSCs (qNSCs) and activated NSCs (aNSCs) have unique autofluorescence profiles. Specifically, qNSCs display an enrichment of autofluorescence localizing to a subset of lysosomes, which can be used as a graded marker of NSC quiescence to predict cell behavior at single-cell resolution. Coupling autofluorescence imaging with single-cell RNA sequencing, we provide resources revealing transcriptional features linked to deep quiescence and rapid NSC activation. Together, we describe an approach for tracking mouse NSC activation state and expand our understanding of adult neurogenesis.

神经干细胞 (NSC) 必须退出静止状态才能产生神经元；然而，我们对这一过程的理解仍然受到当前技术的技术限制的限制。自体荧光代谢辅助因子的荧光寿命成像 (FLIM) 已用于其他细胞类型，以研究代谢重塑驱动的细胞状态变化，从而改变这些内源荧光团的光学特性。使用这种非破坏性、活细胞和无标记策略，我们发现静态 NSC (qNSC) 和激活 NSC (aNSC) 具有独特的自发荧光特征。具体来说，qNSC 显示出局部于溶酶体子集的自发荧光富集，可用作 NSC 静止的分级标记，以单细胞分辨率预测细胞行为。将自发荧光成像与单细胞 RNA 测序相结合，我们提供了揭示与深度静止和快速 NSC 激活相关的转录特征的资源。我们共同描述了一种追踪小鼠 NSC 激活状态的方法，并扩展了我们对成体神经发生的理解。