A High Speed Confocal Microscope for Tracking Single Molecules

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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.