A CONFOCAL MICROSCOPE FOR VISION AND AUDITORY RESEARCH

此申请向美国国立卫生研究院共享仪器拨款计划基金购买共聚焦成像显微镜。来自哈佛医学院眼科和耳鼻喉科的九名研究人员将成为该资源的主要用户群。这个位于马萨诸塞州眼耳医院（MEEI）的小组已经加入努力开发一个新的成像显微镜中心，用于视觉和听觉的结构和功能研究。我们寻求一种最先进的仪器，它能提供非常高的分辨率来成像视网膜或耳蜗组织中的小神经元和支持细胞。我们的工作范围跨越物种，但包括检查免疫荧光标记的细胞，蛋白质或核酸复合物，以及来自携带报告基因构建的转基因动物（果蝇，斑马鱼，小鼠）的组织。仪器必须最大限度地进行低照度和高检测，以尽量减少样品褪色和损坏。此外，我们寻求光学切片分子集合或标记细胞的能力，以生成连续构建的3d图像，以便共定位分子结构并通过切片追踪长神经元。对于核心设施，我们要求光谱共聚焦显微镜，这将提供使用几个荧光团标记的灵活性，并提高准确共定位两个或更多标记结构的能力。所寻求的仪器配置为能够升级额外的激发源或多光子用于未来的应用。为管理、培训和维护该设施提供了一个计划，该计划将在未来几年内由视力和听力小组共享。共聚焦显微镜成像设备为视觉和听力研究的发现带来了新的技术可能性，现在由我们的许多NIH研究基金支持。

This application to the NIH Shared Instrumentation Grants Programs funds to purchase a Confocal Imaging Microscope. Nine research faculty from the Departments of Ophthalmology and Otolaryngology at Harvard Medical School will form the major user group for this resource. This group, located at the Massachusetts Eye and Ear Infirmary (MEEI), has joined in an effort to develop a new imaging microscopy center for structure and function studies in vision and hearing. We seek a state-of-the-art instrument that provides exceptionally high resolution to image small neuronal and support cells in retinal or cochlear tissues. Our work ranges across species, but includes examining immunofluorescently marked cells, protein or nucleic acid complexes, and tissues from transgenic animals (Drosophila, Zebrafish, mice) carrying reporter constructs. The instrument must be maximized for low level illumination and high detection to minimize sample bleaching and damage. In addition, we seek the ability to optically section molecular ensembles or marked cells to generate serially constructed 3-D images in order to co-localized molecular structures and to trace long neurons through sections. For a core facility we request a spectral confocal microscope that will provide flexibility of using several fluorophore markers and enhance the ability to accurately co-localize two or more marked structures. The instrument sought is configured to enable upgrading with additional excitation sources or multi-photon use for future applications. A plan is offered for administration, training, and maintenance of this facility, which will be shared by the vision and hearing groups over the coming years. The confocal microscope imaging facility brings new technical possibilities for discovery in vision and hearing research now supported by our many NIH research grants.