Talamond Pascale, Verdeil Jean-Luc, Conéjéro Geneviève
Institut des Sciences de l'Evolution Montpellier ISE-M, Université Montpellier, CNRS, IRD, EPHE, CC 065, Place Eugène Bataillon, 34095 Montpellier, France.
Histocytology and Plant Cell Imaging platform PHIV, UMR AGAP (CIRAD, INRA, SupAgro)-UMR B&PMP (INRA, CNRS, SupAgro, Montpellier University), 34095 Montpellier, France.
Molecules. 2015 Mar 19;20(3):5024-37. doi: 10.3390/molecules20035024.
Autofluorescent molecules are abundant in plant cells and spectral images offer means for analyzing their spectra, yielding information on their accumulation and function. Based on their fluorescence characteristics, an imaging approach using multiphoton microscopy was designed to assess localization of the endogenous fluorophores in living plant cells. This method, which requires no previous treatment, provides an effective experimental tool for discriminating between multiple naturally-occurring fluorophores in living-tissues. Combined with advanced Linear Unmixing, the spectral analysis extends the possibilities and enables the simultaneous detection of fluorescent molecules reliably separating overlapping emission spectra. However, as with any technology, the possibility for artifactual results does exist. This methodological article presents an overview of the applications of tissular and intra-cellular localization of these intrinsic fluorophores in leaves and fruits (here for coffee and vanilla). This method will provide new opportunities for studying cellular environments and the behavior of endogenous fluorophores in the intracellular environment.
自发荧光分子在植物细胞中大量存在，光谱图像为分析其光谱提供了手段，可得出有关其积累和功能的信息。基于其荧光特性，设计了一种使用多光子显微镜的成像方法来评估活植物细胞中内源性荧光团的定位。该方法无需事先处理，为区分活组织中多种天然存在的荧光团提供了一种有效的实验工具。结合先进的线性解混技术，光谱分析扩展了可能性，并能够可靠地分离重叠发射光谱，同时检测荧光分子。然而，与任何技术一样，确实存在产生人为结果的可能性。这篇方法学文章概述了这些内在荧光团在叶片和果实（此处以咖啡和香草为例）中的组织和细胞内定位的应用。该方法将为研究细胞环境以及内源性荧光团在细胞内环境中的行为提供新的机会。
