One of the remaining challenges for microscopy is an effective and efficient method for imaging three dimensional structures. Existing microscopy methods cannot simultaneously provide both high resolution and high depth-of-field, nor provide z-axis position information without acquiring a serial stack of optical sections at different levels of focus. A novel optical/computational wide-field microscopy method has been developed during Phase I of this project that can image structures in the plane of focus with high resolution, and at the same time image out of focus structures with high depth-of-field. This method also effectively extracts and displays the relative z-axis position information for structure around the plane of focus without having to change the focus of the microscope. In contrast to many other recent developments in microscopy, the simplicity and anticipated cost savings of this method will expand user accessibility. The increased efficiency in photon utilization is anticipated to decrease photobleaching and phototoxicity. Phase I of this project demonstrated feasibility. Phase II seeks to make further improvements in image quality and in efficiency of image acquisition.
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