A major bottleneck limiting understanding of mechanisms and consequences of cell death in complex organisms is the inability to induce and visualize this process with spatial and temporal precision in living animals. Here we report a technique termed two-photon chemical apoptotic targeted ablation (2Phatal) that uses focal illumination with a femtosecond-pulsed laser to bleach a nucleic acid-binding dye causing dose-dependent apoptosis of individual cells without collateral damage. Using 2Phatal, we achieve precise ablation of distinct populations of neurons, glia and pericytes in the mouse brain and in zebrafish. When combined with organelle-targeted fluorescent proteins and biosensors, we uncover previously unrecognized cell-type differences in patterns of apoptosis and associated dynamics of ribosomal disassembly, calcium overload and mitochondrial fission. 2Phatal provides a powerful and rapidly adoptable platform to investigate in vivo functional consequences and neural plasticity following cell death as well as apoptosis, cell clearance and tissue remodelling in diverse organs and species.

中文翻译：

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体内确定细胞类型的靶向双光子化学凋亡消融。

限制对复杂生物中细胞死亡的机制和后果的理解的一个主要瓶颈是无法在活体动物中以时空精度来诱导和可视化这一过程。在这里，我们报告了一种称为双光子化学凋亡靶向消融（2Phatal）的技术，该技术使用飞秒脉冲激光进行聚焦照明来漂白核酸结合染料，从而导致单个细胞的剂量依赖性凋亡而没有附带损害。使用2Phatal，我们可以精确消融小鼠大脑和斑马鱼中不同神经元，神经胶质细胞和周细胞的种群。当与靶向细胞器的荧光蛋白和生物传感器结合使用时，我们发现以前无法识别的细胞类型在凋亡模式和核糖体拆卸相关动力学方面的差异，钙超载和线粒体裂变。2Phatal提供了一个功能强大且可快速采用的平台，可用于研究细胞死亡以及各种器官和物种中的细胞凋亡，细胞清除率和组织重塑后的体内功能后果和神经可塑性。