双光子荧光共振能量转移/荧光寿命成像在脑神经元中的应用。
Two-Photon FRET/FLIM Imaging of Cerebral Neurons.
机构信息
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, ON, Canada.
Department of Molecular Genetics, Faculty of Medicine, University of Toronto, Toronto, ON, Canada.
出版信息
Methods Mol Biol. 2024;2794:33-43. doi: 10.1007/978-1-0716-3810-1_4.
Two-photon FRET (Förster resonance energy transfer) and FLIM (fluorescence lifetime imaging microscopy) enable the detection of FRET changes of fluorescence reporters in deep brain tissues, which provide a valuable approach for monitoring target molecular dynamics and functions. Here, we describe two-photon FRET and FLIM imaging techniques that allow us to visualize endogenous and optogenetically induced cAMP dynamics in living neurons with genetically engineered FRET-based cAMP reporters.
双光子 FRET（Förster 共振能量转移）和 FLIM（荧光寿命成像显微镜）可用于检测深部脑组织中荧光报告分子的 FRET 变化，为监测靶标分子的动态和功能提供了一种有价值的方法。在这里，我们描述了双光子 FRET 和 FLIM 成像技术，这些技术使我们能够使用基于 FRET 的遗传工程 cAMP 报告基因来可视化活神经元中的内源性和光遗传学诱导的 cAMP 动力学。
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