RIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa, Japan.
Department of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Kanagawa, Japan.
Methods Mol Biol. 2022;2525:259-266. doi: 10.1007/978-1-0716-2473-9_19.
Adenosine triphosphate (ATP) is a central metabolite that functions as the energy currency in a living cell. Therefore, visualizing cellular ATP dynamics provides the fundamental information necessary to understand the molecular events involving life phenomena. Live cell imaging technologies using fluorescence (FL)-based indicators have been developed to analyze the dynamics of various biological processes, such as intracellular ATP synthesis and consumption. However, the application of FL-based indicators to plant cells is limited due to the presence of strong chlorophyll autofluorescence, which drastically worsen the signal-to-noise ratio. The bioluminescent (BL) indicators that do not require excitation light could overcome this problem. In this chapter, we introduce a methodology to analyze ATP dynamics in plant cells using BL ATP indicators.
三磷酸腺苷（ATP）是一种重要的代谢物，在活细胞中作为能量货币发挥作用。因此，对细胞内 ATP 动态的可视化可以提供理解涉及生命现象的分子事件所必需的基本信息。利用基于荧光（FL）的指示剂的活细胞成像技术已被开发出来，用于分析各种生物过程的动力学，例如细胞内 ATP 的合成和消耗。然而，由于存在强烈的叶绿素自发荧光，基于 FL 的指示剂在植物细胞中的应用受到限制，这大大降低了信噪比。不需要激发光的生物发光（BL）指示剂可以克服这个问题。在本章中，我们介绍了一种使用 BL ATP 指示剂分析植物细胞内 ATP 动态的方法。
