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Computational adaptive optics for live three-dimensional biological imaging

Summary: Computational adaptive optics for live three-
dimensional biological imaging
Z. Kam*, B. Hanser
, M. G. L. Gustafsson
, D. A. Agard§
, and J. W. Sedat¶
*Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel; and Graduate Group in Biophysics, and Department of
Biochemistry and Biophysics, University of California, San Francisco, CA 94143
Edited by Shinya Inoue´, Marine Biological Laboratory, Woods Hole, MA, and approved December 28, 2000 (received for review June 15, 2000)
Light microscopy of thick biological samples, such as tissues, is
often limited by aberrations caused by refractive index variations
within the sample itself. This problem is particularly severe for live
imaging, a field of great current excitement due to the develop-
ment of inherently fluorescent proteins. We describe a method of
removing such aberrations computationally by mapping the re-
fractive index of the sample using differential interference contrast
microscopy, modeling the aberrations by ray tracing through this
index map, and using space-variant deconvolution to remove
aberrations. This approach will open possibilities to study weakly
labeled molecules in difficult-to-image live specimens.


Source: Agard, David - Department of Biochemistry and Biophysics, University of California at San Francisco


Collections: Biotechnology; Biology and Medicine