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制层面上理解具有特定行为的神经元网络的功能。总的来说，这涉及到局部网络中不同类型神经元之间的解剖连接，这些细胞的生理特性，以及回路中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通性，还在于有效地记录神经元网络在行为过程中的信号活动，从而提供功能信息。自2001年以来，Prairie Technologies, Inc.一直致力于设计、制造、改进和销售一种新型共聚焦扫描系统，即扫场共聚焦显微镜（SFC）。SBIR一期和二期合同为Prairie开发生物市场入门级产品提供了必要的资金。由于成功完成了第一阶段和第二阶段的所有目标，我们已经为公司销售了60多套产品，总销售额为600万美元。最近，我们与尼康仪器公司合作开发了SFC的全球销售渠道。由于速度快、毒性低，SFC在神经和细胞生物学领域越来越受欢迎。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，并增加荧光共振能量转移（FRET）等仪器可以执行的应用数量。特别是，我们希望增加定量技术，如荧光寿命成像（FLIM），用于动态过程的研究和FRET分析。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，而且能够以高分辨率提供局部神经元网络的动态信息。此外，随着该项目的完成，Prairie可以将SFC的产品销售额增加到每年2亿美元的全球激光扫描显微镜市场。

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.