Rapid-Scanning Prairie Multiphoton Microscope System for Molecular Biology Models

描述（由申请人提供）：本申请要求资金购买草原仪器Ultima多光子显微镜系统。这种新仪器将大大扩展普林斯顿大学目前的荧光显微镜能力。Prairie系统将使研究人员能够在高帧率下在标记有多个荧光探针的厚标本中进行多光子显微镜检查，同时减少光损伤并避免光散射进入和离开标本的信号丢失。普林斯顿大学现有的显微镜是传统的共焦显微镜，限制了焦深和可能的观察时间。Prairie系统的多光子帧扫描能力使研究人员能够在散射阻止单光子方法（例如共聚焦显微镜）穿透的组织中获得具有高空间分辨率的动态共定位数据。例子包括苍蝇胚胎和卵巢，发育中的斑马鱼，以及从骨骼到大脑的哺乳动物组织。该系统通过使用声光器件进行光束转向来实现快速帧扫描，从而超过了使用安装在振镜上的振镜所施加的扫描速度限制。与仅通过激发物镜的光收集相比，子级检测将使信号收集增加两倍或更多倍。除了这些成像功能，第二个激光器和一套检流计允许荧光蛋白的光活化和合成“笼”信号分子的光解。总的来说，这些能力将使普林斯顿大学的研究人员能够以电影级的时间分辨率在亚细胞尺度上跟踪快速移动的细胞内颗粒、形态变化和生化动力学。目前在普林斯顿大学进行的研究将大大受益于草原系统，包括果蝇卵母细胞和胚胎中的mRNA运输和细胞运动，神经元细胞中的疱疹病毒组装和运输，完整脑组织中神经胶质细胞和神经元的多细胞信号传导，以及乳腺组织中正常和癌性导管的形态发生。对这些生物系统中的每一个的研究将提供与重大人类健康问题有关的有价值的信息，例如出生缺陷、病毒感染、神经系统疾病和癌症。

DESCRIPTION (provided by applicant): This application requests funds to purchase a Prairie Instruments Ultima multiphoton microscope system. This new instrument will greatly extend the current capabilities for fluorescence microscopy at Princeton University. The Prairie system will allow researchers to perform multiphoton microscopy deep in thick specimens labeled with multiple fluorescent probes at high frame rates, while reducing photodamage and avoiding loss of signal from scattering of light entering and exiting the specimen. Existing microscopes at Princeton University are conventional confocal microscopes, limiting the depth of focus and the duration of observation that is possible. The Prairie system's multiphoton frame scanning capability allows researchers to acquire dynamic colocalization data with high spatial resolution in tissues where scattering prevents one-photon methods (e.g. confocal microscopy) from penetrating. Examples include fly embryos and ovaries, developing zebrafish, and mammalian tissues ranging from bone to brain. The system accomplishes fast frame scanning by using acousto-optical devices for beam steering, thus exceeding the scan speed limit imposed by using galvanometer-mounted mirrors. Substage detection will increase the signal collection by a factor of two or more compared with light collection through the excitation objective alone. In addition to these imaging features, a second laser and set of galvanometers allow photoactivation of fluorescent proteins and photolysis of synthetic "caged" signaling molecules. Overall, these capabilities will allow Princeton researchers to track rapidly moving intracellular particles, morphological change, and biochemical dynamics on a subcellular scale with movie-rate temporal resolution. Research currently undertaken at Princeton which will benefit greatly from the Prairie system include studies of mRNA transport and cell movement in Drosophila oocytes and embryos, herpes virus assembly and transport in neuronal cells, multicellular signaling by glia and neurons in intact brain tissue, and morphogenesis of normal and cancerous ducts in breast tissue. Study of each of these biological systems will provide valuable information pertaining to significant human health problems such as birth defects, viral infection, neurological disease, and cancer.