Two-Photon Confocol Microscope for Biological Imaging in Plants

多光子共聚焦显微镜的采集。David W. Galbraith：首席研究员。摘要：由美国国家科学基金会生物仪器项目资助购买一台BioRad mrc - 10242p多光子共聚焦显微镜。该显微镜将主要服务于15名教师用户，他们每个人都有活跃的联邦研究资助，其中大部分来自NSF，总计约2500万美元。它将以多用户设施的形式在亚利桑那大学农业学院提供。这台双光子显微镜是亚利桑那州的第一台同类产品，因此也将向亚利桑那大学和该州其他机构的所有感兴趣的用户开放。与传统的共聚焦显微镜相比，双光子显微镜具有显著的优点。特别是，它允许在最大限度地减少组织损伤的条件下分析活组织，提高光的穿透深度，并允许同时激发多个荧光标记。将受益于获得显微镜的重点项目包括：(a)分析植物对施加的离子和干旱胁迫的反应。该项目旨在全面描述参与这一过程的基因和基因产物，并将这些基因转移到植物中以提高对这些胁迫的抗性/耐受性。(b)玉米表达基因的功能分析。该项目旨在对玉米植株生命周期中表达的全部基因进行编目。这一信息将导致玉米作为一种作物的改进。(c)细胞色素P450基因超家族在植物中的功能分析。该项目的重点是发现每个P450基因的酶功能（预计在拟南芥中超过340个）。这些酶催化产生多种重要的植物次生化合物的途径，具有杀虫剂和抗癌剂等价值。在所有情况下，双光子显微镜将影响这些项目，因为它能够快速分析基因表达的特征，这是使用其他仪器无法实现的。特别是，它使科学家能够确定给定基因在何时何地开启和关闭，以及由这些基因表达产生的蛋白质在细胞和组织中的位置。

Acquisition of a multiphoton confocal microscope. David W. Galbraith: Principal Investigator. Abstract:Funding from the NSF Biological Instrumentation Program will be employed to purchase a BioRad MRC-1024 2P multi-photon confocal microscope. The microscope will primarily service 15 faculty users each of whom have active federal research grants, the majority from NSF, totalling about $25,000,000. It will be made available in the form of a multi-user facility in the College of Agriculture at the University of Arizona. The two-photon microscope is the first of its kind in the state of Arizona, and therefore will also be made available to all interested users at the University of Arizona and those of other institutions within the state.The two-photon microscope possesses significant advantages over conventional confocal microscopy. In particular, it allows analysis of living tissues under conditions that minimize tissue damage, improve the depth of penetration of the light, and allow simultaneous excitation of multiple fluorescent labels. Highlighted projects that will benefit from acquisition of the microscope include: (a) The analysis of the responses of plants to imposed ionic and drought stress. This project aims to provide a comprehensive description of the genes and gene products that are involved in this process, and the transfer of these genes into plants for improved resistance/tolerance to these stresses. (b) The functional analysis of expressed genes in maize. This project aims to catalog the totality of genes that are expressed during the life cycle of the maize plant. This information should lead to improvement of maize as a crop plant. (c) The analysis of the function of the cytochrome P450 gene superfamily in plants. This project centers on discovering the enzymatic functions of each of the individual P450 genes (over 340 are expected in Arabidopsis thaliana). These enzymes catalyze pathways leading to a variety of important plant secondary compounds, of value as pesticides and as anti-cancer agents, amongst other things. In all cases, the two-photon microscope will impact these projects since it enables rapid analysis of features of gene expression that cannot be achieved using other instrumentation. In particular, it enables the scientist to determine where and when given genes are turned on and off, and where the proteins produced from expression of these genes are located within cells and tissues.