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Summary: Practical Image Restoration of Thick Biological Specimens Using Multiple
Focus Levels in Transmission Electron Microscopy
Karen F. Han,1 John W. Sedat, and David A. Agard
Graduate Group in Biophysics, Department of Biochemistry and Biophysics, Howard Hughes Medical Institute,
University of California Medical School, San Francisco, California 94143-0448
Received September 9, 1997
Three-dimensional electron tomographic studies
of thick specimens such as cellular organelles or
supramolecular structures require accurate inter-
pretations of transmission electron micrograph in-
tensities. In addition to microscope lens aberra-
tions, thick specimen imaging is complicated by
additional distortions resulting from multiple elas-
tic and inelastic scattering. Extensive analysis of
the mechanism of image formation using electron
energy-loss spectroscopy and imaging as well as exit
wavefront reconstruction demonstrated that mul-
tiple scattering does not contribute to the coherent
component of the exit wave (Han et al., 1996, 1995).
Although exit wavefront restored images showed
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