A Confocal Laser Scanning Microscope for Neuroscience Imaging Center

描述（由申请人提供）：该提案是为田纳西大学健康科学中心的神经科学成像中心购买一台具有宽光谱扫描能力，高分辨率和高灵敏度的现代共聚焦激光共聚焦扫描显微镜（蔡司LSM 710）。LSM将安装在蔡司Axio观察者倒置显微镜上，在检查材料类型方面提供很大的灵活性。显微镜台将有几个适配器，这样活的神经元可以在各种孵育井的温暖，充氧，潮湿的环境中成像，或者通过切换适配器，固定的脑切片可以用荧光探针标记的特定结构和/或分子成像。LSM配备了6条激光线和3个光电倍增管（pmt），能够扫描34个通道，并且比以前的LSM更敏感。这种灵敏度意味着需要更少的激光功率，因为激光功率会损害活细胞，并使这些研究中使用的探针漂白。这些荧光探针可以跨越各种发射波长，并且多达10个探针可以同时被观察和表征。光谱扫描灵敏度使得具有10 nm或更小荧光发射谱线的探针可以被分离、图形标记和分析。因此，神经科学家将有很大的灵活性来确定神经元的精确位置、生化性质、它们的广泛过程和突触连接。该LSM的空间分辨率（6144 x 6144）与蔡司物镜和共聚焦光学相结合，意味着可以观察、测量和计数远低于传统光学显微镜（~0.10 m）极限的结构。这里的神经科学家将研究一系列广泛的问题，从神经元如何形成适当的连接，到了解哪些离子通道塑造了通信所必需的电行为。这些信息对于理解神经退行性或神经性疾病如帕金森氏症、亨廷顿氏症和肌张力障碍是如何产生和发展的至关重要，对于理解我们如何治疗这些疾病至关重要。在各种遗传条件下异常表达的脑蛋白，例如与某些形式的自闭症有关的那些，也将用这台显微镜进行检查。实时成像能力意味着可以测试药物在对抗脑肿瘤细胞方面的功效，并获得实时结果。许多大脑区域的基本结构，如嗅觉、脑垂体激素的神经系统控制、感觉-运动整合、运动和认知的协调，以及正常的大脑发育和乙醇的有害影响，都将被检查。

DESCRIPTION (provided by applicant): The proposal is to purchase a modern confocal laser confocal scanning microscope (Zeiss LSM 710) with wide spectral scanning capability, high resolution, and high sensitivity for a Neuroscience Imaging Center at the University of Tennessee Health Science Center. The LSM will be mounted on a Zeiss Axio Observer inverted microscope, providing great flexibility in the type of material examined. The microscope stage will have several adaptors such that live neurons may be imaged in a warmed, oxygenated, humidified environment in a variety of incubation wells, or by switching adapters, fixed brain slices may be imaged with specific structures and/or molecules labeled with fluorescent probes. The LSM comes equipped with six laser lines, and three photomultipliers (PMTs) capable of scanning 34 channels, and is much more sensitive than previous LSMs. This sensitivity means that less laser power, which is damaging to live cells and which bleaches the probes used in these studies, is required. These fluorescent probes can span a variety of emitted wavelengths, and up to 10 probes may be simultaneously viewed and characterized. The spectral scanning sensitivity is such that probes with fluorescent emission profiles 10 nm or even less may be separated, graphically marked, and analyzed. Thus, neuroscientists will have great flexibility to determine the precise location, and biochemical nature, of neurons, their extensive processes, and their synaptic connections. The spatial resolution of this LSM (6144 x 6144) combined with Zeiss objectives and confocal optics, means that structures well below the limit of conventional light microscopes (~0.10 5m) may be viewed, measured and counted. Neuroscientists here will be studying a broad array of problems, from how neurons develop appropriate connections, to understanding which ion channels sculpt their electrical behavior necessary for communication. This information is critical to understanding how neurodegenerative or neuropathic diseases such Parkinson's, Huntington's, and dystonia arise and progress - information critical to understanding how we might cure these conditions. Brain proteins abnormally expressed in a variety of genetic conditions, such as those related to some forms of autism, also will be examined with this microscope. Live imaging capability means that drugs may be tested for their efficacy in fighting brain tumor cells, with real-time results. The basic structures of many brain areas, such as those serving olfaction, nervous system control of pituitary hormones, sensory-motor integration, coordination of movement and cognition will be examined, as will normal brain development, and the deleterious effects of ethanol.