The Swept Field Confocal Microscope

描述（由申请人提供）：神经科学的主要目标是在机械水平上理解具有特定行为的神经元网络的功能。这通常涉及局部网络内各种类型神经元之间的解剖学连接性、这些细胞的生理特性以及电路内神经元的时空放电信号。挑战不仅在于发现神经元之间的连接，而且还在于以提供功能信息的方式有效地记录行为期间神经元网络内的信号活动。自2001年以来，Prairie Technologies，Inc.一直在设计、制造、改进和销售一种新型的共焦扫描系统-扫掠场共焦显微镜（SFC）。SBIR第一阶段和第二阶段奖项为Prairie开发生物市场入门级产品提供了必要的资金。由于成功完成了我们所有的第一阶段和第二阶段的目标，我们已经能够销售超过60个单位，总销售额为600万美元的公司。最近，我们与尼康仪器公司合作，为SFC开发了一个全球销售渠道。由于SFC的高速度和低毒性，现在在神经和细胞生物学领域的销售普及。为了提供更多的功能信息，我们建议进一步提高SFC的速度，简化其对科学家的易用性，并增加可以使用仪器（如荧光共振能量转移（FRET））进行的应用数量。特别是，我们想增加定量技术，如荧光寿命成像（FLIM）的动态过程和FRET分析的研究。通过完成本提案中概述的具体目标，SFC不仅可以作为共聚焦显微镜，而且有能力提供有关局部神经元网络的高分辨率动态信息。此外，随着该项目的完成，Prairie可以增加SFC的产品销售额，进入每年2亿美元的全球激光扫描显微镜市场。 公共卫生相关性：神经科学的一个主要目标是在机械水平上理解具有特定行为的神经元网络的功能，这可以用来表征神经系统疾病。这些研究需要能够识别信号网络并非侵入性地研究活细胞内的动力学。改进的扫描场共聚焦显微镜系统将提供这样的能力，以记录正常和患病过程的真实的时间的时空信息，并研究细胞动力学与荧光寿命成像检测到的微环境和信号通路的额外信息。

DESCRIPTION (provided by applicant): A primary goal in neuroscience is to understand at a mechanistic level the functioning of neuronal networks with specific behaviors. This involves in general, the anatomical connectivity among the various types of neurons within the local network, the physiological properties of these cells, and the spatiotemporal firing signals of the neurons within the circuit. The challenge is not only to discover the connectivity among neurons but also to effectively record the signal activity within the neuronal network during a behavior in such a way as to provide functional information. Since 2001 Prairie Technologies, Inc. has been designing, building, refining and selling a new type of confocal scanning system, the Swept Field Confocal microscope (SFC). SBIR Phase I and II awards have provided the funding necessary for Prairie to develop an entry level product for the biological marketplace. Due to successful completion of all of our Phase I and Phase II goals, we have been able to sell over 60 units totaling to sales of $6,000,000.00 for the company. Recently we have been able to develop a worldwide sales channel for the SFC with Nikon Instruments, Inc. Because of high speed and low toxicity, the SFC is now gaining in sales popularity in the neuro and cell biology fields. In order to provide more functional information, we propose to further improve the speed of the SFC and simplify its ease of use for the scientists and to increase the amount of applications that can be performed with the instrument such as Fluorescence Resonance Energy Transfer (FRET). In particular, we would like to add quantitative techniques such as Fluorescence Lifetime Imaging (FLIM) for the study of dynamic processes and FRET analysis. By completing the specific aims outlined in this proposal, the SFC can not only perform as a confocal microscope, but have capability to provide dynamic information about the local neuron network with high resolution. In addition, with completion of this project Prairie can achieve increased product sales of the SFC into the $200 million annual world-wide laser scanning microscopy market. PUBLIC HEALTH RELEVANCE: A primary goal in neuroscience is to understand at a mechanistic level the functioning of neuronal networks with specific behaviors, which can be used to characterize neurological disorders. Such studies require the ability to identify the signaling networks and noninvasively study the dynamics within living cells. The improved Swept Field Confocal microscope system will provide such capabilities to record the spatiotemporal information in real time of normal and diseased processes and to study the cellular dynamics with additional information about the microenvironment and signaling pathways detected with Fluorescent Lifetime Imaging.