A New Generation, FAST 3D Fluorescence Microscope Using Wavefront Coded Optics

Authors:
Carol Cogswell, Matthew Arnison, David Philp: School of Physics, University of Sydney, NSW2006 Australia

Edward Dowski, Sara Tucker, W. Thomas Cathey: Dept. of Electrical and Computer Engineering, University of Colorado, Boulder CO 80309

Summary:

We are two research groups that are collaborating to develop a "new-generation" fluorescence microscope that will allow very fast (milliseconds) acquisition of "extended-depth-of-focus" images from live-cell biological applications. Our new microscope system incorporates the innovative techniques of wavefront coding pioneered by two of the authors (W.T. Cathey and E. Dowski) at the University of Colorado. These pages represent some of our preliminary results that were recently presented (as posters) at the following three conferences/workshops:

          * Microscopy & Microanalysis '99 (Microscopical Society of America Conference), 1-5
              August 1999, Portland, OR
          * NIH Workshop on In Vivo Optical Imaging, 16-17 September 1999, Bethesda, MD
          * 1999 Optical Society of America Annual Meeting, 26-30 September 1999, Santa Clara,
              CA


The Problem:

For 3D real-time fluorescence imaging of live-cell dynamics and in vivo processes, all confocal and widefield (deconvolution) microscopes are too slow because they require sequential acquisition of many planes of focus to build up a 3D image.


Our Solution:

We are at the "beginning stages" of developing a totally different approach to 3D fluorescence microscopy that promises to be much faster than contemporary confocal or widefield techniques.

Instead of plane-by-plane image acquisition being required to remove out-of-focus blurring, our new system produces an extended-depth-of-focus (EDF) image. It does so with a single exposure on an ICCD camera, followed by a single-iteration, digital deconvolution filtering step that is currently fast enough to keep up with a CCD running at 15-30 frames/sec.

How it works - diagrams and explainations
Our Results - new EDF fluorescence images
Other Selected Publications



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