"IDBR: TYPE A" A fluorescence microscope optical insert that creates video-rate 3D imaging capabilities using an innovative "expanded point information content" design

University Of Colorado At Boulder, Boulder CO

Investigators

Abstract

Non Technical Description: Scientists at the University of Colorado at Boulder will develop a modified biological microscope whose purpose is to demonstrate how a novel approach to optical design, called expanded point information content (EPIC), can convert existing live-cell fluorescence microscopes into fully 3D video-rate imaging instruments at a fraction of the cost and complexity of commercial confocal designs. The new microscope will demonstrate to biologists engaged in studies of live cell dynamics that it will be possible to record high resolution 3D images at speeds an order of magnitude faster than existing microscopes and simultaneously locate features in depth to an accuracy of 70nm or better, without the need to change focus. The project is grounded in the field of computational optics and is based on the premise that, by optically altering the microscope point spread function, much more information about the original three-dimensional sample can be encoded into pairs of simultaneously recorded images and then rapidly retrieved by digital image processing. If this procedure (known as point spread function engineering) is done in a controlled manner, the expanded information from each 3D object point, once processed, will provide biologists with the capability to obtain 3D animations of subcellular dynamic events that occur at video rates or faster. Technical Description: In a broader context, the new EPIC approach represents a potentially transformative concept that could revolutionize the way next generation optical imaging instruments are designed, not only in microscopy, but also in other devices such as endoscopes and telescopes. Implementation of the EPIC design requires only simple modifications to a standard optical imaging system, such as inserting a phase plate, adding a second camera and installing a software package. In this project, these modifications will be implemented on a commercial biological microscope. Once completed, the new microscope will be tested by collaborating biologists in challenging studies of living cells. It will also provide the basis for teaching and training young scientists and engineers, providing opportunities for conducting multi-disciplinary training workshops where biology students learn about optical microscopy and image processing, and engineering students learn about challenges in live-cell dynamics investigations. A final planned activity will be to work with collaborating commercial partners to bring the new EPIC microscope add-on system to the marketplace, thereby making it readily accessible to the many hundreds of biologists who use fluorescence microscopes as a primary laboratory tool.