A High Speed Confocal Microscope for Tracking Single Molecules

该奖项支持将高速成像技术的最新进展应用于共焦显微镜。这项工作的目标是开发一种旋转圆盘共聚焦显微镜，具有所需的速度和灵敏度，以便能够成像和跟踪单个荧光标记的蛋白质在三维中快速扩散。亚细胞过程的光学成像是细胞生物学中的一个重要工具，但涉及少量蛋白质的快速动态过程很难或不可能用现有的仪器可视化。开发工作在系统集成、图像处理和数据处理方面面临重大挑战。性能目标包括通过快速z扫描以高达每秒1000帧的速度获取高分辨率共焦图像，以便以高速率进行全三维图像堆叠。将开发集成的软件和硬件，用于接近基本探测极限的成像，以及处理和处理将产生的海量数据。该仪器的能力将通过成像以前无法观察到的细胞骨架网络的动态、轴突的生长和发育以及其他基本的细胞生物学过程来测试。作为努力的一部分，研究生和博士后研究员将接受有关仪器开发和软件开发方面的培训。

This award provides support for the application of recent advances in high speed imaging technology to confocal microscopy. The goal of the effort is the development of a spinning disk confocal microscope with the combination of speed and sensitivity required to enable imaging and tracking of individual fluorescently labeled proteins rapidly diffusing in three dimensions. Optical imaging of subcellular processes is a critical tool in cell biology, but rapid dynamical processes involving small numbers of proteins are difficult or impossible to visualize using existing instruments. The development effort faces significant challenges for systems integration, image processing, and data handling. Performance goals include the acquisition of high resolution confocal images at up 1000 frames per second with fast z-scanning in order to allow for full 3-dimensional image stacks at high rates. Integrated software and hardware will be developed for imaging near the fundamental detection limits, and for handling and processing the prodigious amounts of data to be generated. The capabilities of the instrument will be tested by imaging previously unobservable dynamics in cytoskeletal networks, growth and development of axons, and other fundamental cell biological processes. As part of the effort, graduate students and postdoctoral fellows will be trained in relevant aspects of instrument development and software development.