MULTISPECTRAL FP-TIR MICROSCOPY FOR NEUROBIOLOGY

该第一阶段SBIR申请是为了响应NIMH SBIR PA- 99-007而提交的，旨在通过结合Dan Axelrod开发的全内反射（TIR）和荧光偏振技术来创建一种新型的显微镜平台（密歇根州）与专有的液晶可调滤波器和偏振设备开发的CRI，沿着与创新的成像算法，以产生具有理想地适合于研究神经生物学中的问题的特征的近视频速率系统。优势：TIR提供了精致的空间分辨率（约100 nm）和独特的偏振态。 荧光偏振可以确定标记的膜和其他结构的平移、旋转或取向，以及小配体的物理状态。 光谱成像赋予灵活性，在使用多种荧光信号和FRET实验。 没有类似的系统可以提供这三个主要功能。目标：（a）建立一个随时可用的TIR荧光偏振显微镜系统。(b)纳入专有CRI，Inc.该技术包括偏振光学器件、可调滤波器和创新的采集和处理算法。(c)开发初步的近视频速率控制和采集软件，用于可行性测试。 可行性：与神经生物学家罗恩霍尔茨（密歇根州）合作：（a）以近视频速率可视化与嗜铬细胞分泌相关的膜折叠和融合事件，（B）在胞吐过程中对单个GFP标记的胞吐颗粒和细胞骨架元件进行成像。拟定商业应用：在这个项目中开发的成像系统的主要市场将是神经生物学，药理学，药物发现和细胞生物学的研究人员。 潜在的大的应用也存在于光谱核型分析的多光谱TIR为基础的成像仪，可以区分10个或更多的探针同时。

This Phase-I SBIR application, submitted in response to NIMH SBIR PA- 99-007, seeks to create a novel microscopy platform by combining total internal reflection (TIR) and fluorescent polarization techniques developed by Dan Axelrod (Michigan) with proprietary liquid crystal tunable filter and polarization devices developed at CRI, along with innovative imaging algorithms, to yield a near-video rate system with features ideally suited to investigating questions in neurobiology. Advantages: TIR provides exquisite spatial resolution (approximately 100 nm) and unique polarization states. Fluorescence polarization can determine translation, rotation, or orientation of labeled membranes and other structures, and the physical state of small ligands. Spectral imaging confers flexibility in the use of multiple fluorescent signals and FRET experiments. No similar system is available that offers these three major features. Goals: (a) Create a ready-to-use TIR-fluorescence polarization microscope system. (b) Incorporate proprietary CRI, Inc. technology, comprising polarization optics, tunable filters and innovative acquisition and processing algorithms. (c) Develop preliminary near-video rate control and acquisition software for sue in feasibility testing. Feasibility: In collaboration with neurobiologist Ron Holz (Michigan): (a) visualize membrane-folding and fusion events related to secretion in chromaffin cells at near-video rate, and (b) image individual GFP-labeled exocytotic granules and cytoskeletal elements during exocytosis. PROPOSED COMMERCIAL APPLICATION: The primary market for the imaging system developed in this project would be to researchers in neurobiology, pharmacology, drug discovery, and cell biology. Potentially large applications also exist in spectral karyotyping for a multi-spectral TIR-based imager that can distinguish 10 or more probes simultaneously.