Digital Imaging Microscope

本提案要求支持一项多学科的努力，以开发用于分析和控制细胞功能基础上分子分布变化的数字成像显微镜。为了提高荧光成像的时空分辨率，我们建议开发新的毫秒级成像显微镜和相机系统，并开发进一步扩展分辨率的计算方法。我们将开发定量分析2-D， 3-D和3-D时间序列荧光图像的方法，并确定在一个隔间中给定分子的浓度估计的误差界限。我们将通过集成图像显示工具和自动特征提取算法的交互，为图像的测量和分析提供强大的能力。我们将开发工具来研究两个或多个分子之间的相互关系。我们将提高算法的计算速度，使数字成像显微镜成为一个实时设备，允许交互或自动控制实验。最后，我们将增加仪器的能力，以控制和修改分子分布在活细胞通过控制光解“笼”化合物。

This proposal requests support for a multidisciplinary effort to develop the digital imaging microscope for the anlaysis and control of changes in molecular distribution underlying cell function. We propose to enhance the spatial and temporal resolution of fluorescence imaging by developing new microscope and camera systems for imaging in the millisecond domain and by developing computational methods for further extending resolution. We will develop methodology for quantitative analysis of 2-D, 3-D and 3-D time series fluorescence images and for determining error bounds on estimates of the concentration of a given molecule in a compartment. We will provide a powerful capability for the measurement and analysis of images by integrating tools for image display and interaction with automated feature extraction algorithms. We will develop tools to investigate the relationship of two or more molecules with respect to one another. We will incrased the algorithm computation speed to make the digital imaging microscope into a real- time device that allows interactive or automated control of experiments. Finally, we will add to the instrument the ability to control and modify molecular distribution in living cells by controlled photolysis of "caged" compunds.