VIDEO RATE CCD RATIO IMAGING CONFOCAL MICROSCOPE

比率荧光成像是一种重要的调查工具 活细胞的生物学功能。共聚焦显微镜允许 细胞功能和结构的三维探索。这 项目将开发一种新型的光学附件，它将允许视频- 在洞察线扫描共焦显微镜上进行速率CCD比率成像， 由Meridian Instruments，Inc.针对给定的帧周期(即 33毫秒)和探测器灵敏度级别，行扫描共焦允许 比点扫描仪更高的信噪比，因为它可以检测到 沿线所有点同时发出荧光，增强 帧期间允许的驻留时间。这一优势使得 生物结构的实时共聚焦成像 点扫描仪的荧光限制。在第一阶段，一种光学 将开发一种分离和过滤两种荧光的设备 来自洞察力图像的波长，并将它们并排传送到 一台ccd摄像机。将选择合适的摄像头和软件 可实现每秒30个或更多比例的图像(488 x 256像素)。原型仪器将允许探索真实的- 时间、共焦比成像以及实时共焦 生物结构的本地化研究。在第二阶段，一个完整的 将开发集成实时比率共焦显微镜。 建议的商业应用： 视频率比成像在洞察力共焦上的可用性 显微镜将极大地拓宽活细胞的应用范围 这个仪器可以用来做什么。这些新的应用，如 因为原位定量的钙和pH测量将提供 研究人员购买我们的共焦显微镜的令人信服的理由。

Ratiometric fluorescence imaging is an important tool for investigation of biological function in living cells. Confocal microscopy allows three-dimensional exploration of cell function and structure. This project will develop a novel optical attachment that will permit video- rate CCD ratio imaging on the InSight line-scanning confocal microscope, offered by Meridian Instruments, Inc. For a given frame period (i.e. 33 msec) and detector sensitivity level, a line-scanning confocal allows a higher signal to noise ratio than a point scanner, because it detects fluorescence at all points along the line at the same time, increasing the dwell time allowed during the frame period. This advantage allows real-time confocal-imaging of biological structures that are often fluorescence-limiting for a point scanner. In Phase I, an optical apparatus will be developed to split and filter two fluorescent wavelengths from the InSight image, and to deliver them side-by-side to a CCD camera. Appropriate cameras will be selected and software developed to accomplish 30, or more, ratioed images per second (488 x 256 pixels). The prototype instrument will allow exploration of real- time, confocal ratio imaging as well as real-time co- localization studies of biological structures. In Phase II, a fully integrated real-time ratio confocal microscope will be developed. PROPOSED COMMERCIAL APPLICATION: The availability of a video-rate ratio imaging on the InSight confocal microscope would greatly broaden the number of live cell applications for which the instrument could be used. These new applications, such as in situ quantitative calcium and pH measurements, would provide compelling reasons for researchers to purchase our confocal microscope.