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Summary: Three-Dimensional Structural Characterization of
Centrosomes from Early DrosophilaEmbryos
Michelle Moritz,* Michael B. Braunfeld,* Jennifer C. Fung,* John W. Sedat,* Bruce M. Alberts, ~
and David A. Agard*
Department of Biochemistry & Biophysics and *Howard Hughes Medical Institute, University of California at San Francisco,
San Francisco, CA 94143-0448
Abstract. An understanding of the mechanism and
structure of microtubule (MT)-nucleating sites within
the pericentriolar material (PCM) of the centrosome
has been elusive. This is partly due to the difficulty in
obtaining large quantities of centrosomes for analysis,
as well as to the problem of attaining interpretable
structural data with conventional EM techniques. We
describe a protocol for isolating a large quantity of
functional centrosomes from early Drosophilaem-
bryos. Using automated electron tomography, we have
begun a three-dimensional structural characterization
of these intact centrosomes with and without regrown
MTs. Reconstructions of the centrosomes to ~6-8 nm
resolution revealed no large structures at the minus
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