Acquisition of a Photomicroscope & Image Analysis System for Plant Biology Research

要求拨款购买一台配备有能够探测弱光发射的摄像机的显微显微镜、一套图像处理和分析系统（用于提高图像质量、数据量化和植物组织三维重建）和一台录像机。该仪器的主要用途之一是分析植物发育过程中的基因表达和蛋白质定位。特别是，我们将继续研究胚胎发生过程中细胞表面蛋白的调控（Grimes博士），植物草酸钙沉积的细胞基础（Franceschi博士），以及参与C4光合作用机制进化的关键酶的时空分布（Ku博士）。在这些研究中，我们将需要分析低水平细胞免疫荧光数据，定量特定细胞和组织类型中的基因表达，并进行图像分析和三维重建，以便全面了解这些过程所涉及的机制。该仪器还将用于通过监测荧光染料在汇组织中的运动来探索韧皮部卸载的机制。(Fisher博士)。为了充分理解韧皮部功能，有必要在结构上下文中评估数据。从相关组织的切片中使用三维重建将对该领域的进展至关重要。苹果酸积累到CAM植物液泡中的机制也正在探索中（Uribe博士）。利用荧光探针监测液泡膜在体内的能量学，将使活的植物细胞内部的事件可视化，而不是研究孤立的膜组分。所要求的仪器对拟议研究的许多方面至关重要，并将大大提高我们在植物细胞生物学研究的能力。

Funds are requested for the purchase of a photomicroscope equipped with a video camera capable of detecting low light emissions, an image processing and analysis system (for enhancement of image quality, quantification of data, and 3-dimensional reconstruction of plant tissues), and a video recorder. One of the primary uses of this instrumentation will be in the analysis of gene expression and protein localization during plant development. In particular, we will pursue investigations into the regulation of cell surface proteins during embryogenesis (Dr. Grimes), the cellular basis of calcium oxalate deposition in plants (Dr. Franceschi), and the temporal and spatial distribution of key enzymes involved in the evolution of the C4 photosysthesis mechanism (Dr. Ku). In these studies, we will need to analyze low-level cell immunofluorescence data, quantitate gene expression in specific cell and tissue types, and perform image analysis and 3-dimensional reconstruction in order to develop a comprehensive understanding of the mechanisms involved in these processes. This instrument will also be used to probe the mechanisms of phloem unloading by in vivo monitoring of fluorescent dye movement in sink tissues. (Dr. Fisher). To fully understand phloem function, it is necessary to evaluate the data in a structural context. The use of 3-dimensional reconstruction from sections of the tissues involved will be of central importance for progress in this area. The mechanism of malic acid accumulation into the vacuole of CAM plants is also being explored (Dr. Uribe). The use of fluorescent probes to monitor the energetics of the the vacuolar membrane in vivo will allow events inside of a living plant cell to be visualized, as opposed to studies on isolated membrane fractions. The requested instrumentation is critical to many aspects of the proposed research and will greatly enhance our capabilities for research in plant cell biology.