DEVELOPMENT OF A HOLOGRAPHIC FLUORESCENCE MICROSCOPE

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This award supports the development of a new kind of fluorescent microscope. This microscope, dubbed the Holoskop, when developed, will allow the viewing of microscopic specimens in 3 dimensions (3D) in real-time.In the last decade, advances in cell science have made it apparent that there is rapid intracellular movement of small molecules and proteins. However there is currently no microscope available to view these events in real time. Green fluorescent proteins obtained from jellyfish and other aquatic organisms (GFP) have been isolated and sequenced. Thus GFP can be expressed in cells as part of the expression of proteins of biological interest by linking the DNA from GFP with the DNA of the protein of interest. Such expression enables one to follow the movement of target proteins of biological importance by following the fluorescence of the GFP tag.These fluorescent protein construct probes, and other organic fluorescent probes, combined with advances in optical sectioning utilizing another type of microscopy called confocal microscopy, have revolutionized studies in 3D localization and dynamic trafficking of proteins, ions, and messenger molecules. In practice, the 3D imaging of probes is derived from stacks of images taken through the z-plane in cells or tissues. Thus fluorescent molecules identifying cells, structures or processes, which move rapidly in the x-y or x-y-z plane during the capture of the image stack, cannot at present be adequately resolved or studied. Knowledge of these fast processes and their interactions is of paramount importance to understanding the myriad of events in the normal and abnormal functioning of cells and tissues. The objective of this project is to develop a new and demonstrably faster microscope for 3D imaging. The Holoskop will utilize scanning holographic principles for accelerated 3D imaging without the need for collection of image stacks.In addition to developing an important new tool for biology, this project will have considerable impact upon the educational and outreach activities inherent in the universities involved in this project. The Holoskop once developed, will aid scores of projects in a wide variety of biological science investigations since there is currently no method that can obtain the kind of information expected from this instrument.