Chanoca Alexandra, Burkel Brian, Grotewold Erich, Eliceiri Kevin W, Otegui Marisa S
Department of Botany, University of Wisconsin-Madison, Madison, WI, USA.
Laboratory of Molecular and Cellular Biology, University of Wisconsin-Madison, Madison, WI, USA.
Methods Mol Biol. 2018;1789:131-141. doi: 10.1007/978-1-4939-7856-4_10.
Anthocyanins are intrinsically fluorescent pigments that accumulate in plant vacuoles. We have developed a platform to analyze the fluorescence decay of anthocyanins by fluorescence lifetime imaging microscopy (FLIM), under in vitro and in vivo conditions. Fluorescence lifetime of a fluorophore can be influenced by temperature, pH, oxygen concentration, and other environmental conditions. Within plant cells, the anthocyanin fluorescence lifetime correlates with distinct subcellular compartments. Vacuolar anthocyanins exhibit shorter fluorescence lifetime than the cytoplasmic pool. Consistent with these observations, lower pH of anthocyanins solutions correlated with shorter fluorescence lifetimes. We discuss here the use of FLIM as a tool for analyzing the subcellular distribution of anthocyanins and estimating variation in vacuolar pH in intact cells.
花青素是积累在植物液泡中的固有荧光色素。我们开发了一个平台，通过荧光寿命成像显微镜（FLIM）在体外和体内条件下分析花青素的荧光衰减。荧光团的荧光寿命会受到温度、pH值、氧气浓度和其他环境条件的影响。在植物细胞内，花青素的荧光寿命与不同的亚细胞区室相关。液泡中的花青素比细胞质中的花青素荧光寿命短。与这些观察结果一致，花青素溶液较低的pH值与较短的荧光寿命相关。我们在此讨论使用FLIM作为分析花青素亚细胞分布和估计完整细胞液泡pH值变化的工具。
