We propose to design and develop a new type of fluorescence microscope, based on our recently-invented Wavefront Coding imaging technology, that will allow biologists to observe living cell dynamics in ways that are not possible with any current microscope system. Wavefront Coding will allow standard fluorescence microscopes to image the entire volume of a specimen with a very large depth of field and with very fast (milliseconds) acquisition rates. Thus, biologists can observe a new spectrum of liver-cell processes. With Wavefront Coding to greatly extended the depth of field (>10X the normal depth of field), cells with substantial thickness can be presented quickly and clearly in a single image. We propose to test the utility of Wavefront Code microscopes in laboratories at the University of Colorado and the National Cancer Institute's Imaging Facility. Particular research at the University of Colorado laboratory to be targeted includes cell biology of fertilization in plants. The first research at the National Cancer Institute's Imaging Facility that will explore the benefits of Wavefront Coding includes viewing motion of PML bodies in promyoleucitic leukemia, steroid hormone receptor function in living cells, and studies of fixed yeast cells containing multiple centrosomes. Our new fluorescence system should also provide significant improvement over existing microscopes in most applications where studies of the dynamics of living cells and their components are of primary interest.
Low-cost and high-speed alternative to confocal and deconvolution systems for forming large depth of field images. Systems can be retrofit to existing microscopes by use of new objectives and processing software, or as a component of new microscopes.
