Biomedical
Imaging
Image Analysis of the Mitotic Spindle
The
Mitotic Spindle
 The mitotic
spindle (see Fig. 1) is a temporary and
critical nuclear structure that serves as the machinery for chromosome
segregation during cell division. The tubulin-based structure of
the spindle is also the target of many anti-cancer drugs. Furthermore,
there are many proteins that interact with the spindle in unknown
ways. In this effort, automated image analysis tools were developed
to quantitatively characterize the mitotic spindle under a variety
of conditions including gene knockout and drug treatment. These
image analysis tools will allow life science researchers to explore
the roles of many proteins in cell cycle progression, spindle checkpoint,
and/or chromosome segregation and, thereby, aid in the discovery
of new drug targets.
Technology
Five cell types were established for study, including wild-type,
two different gene knockouts, and drug-treatment at two different
concentrations.  The cells were fixed and probed with fluorescent-labeled,
anti-tubulin antibodies that attach to the spindles of metaphase
cells. The spindles were then imaged via confocal microscopy, resulting
in a three-dimensional (3D) stack of images for each spindle.
Image
processing tools were developed to extract a set of quantitative
features from each 3D spindle image stack. These features are
based on the spindle's intrinsic coordinate system, which is
referenced to the spindle center of mass and principal axes of
inertia (see Fig 2). Statistical analysis of these features across
all the image stacks in the database has indicated that significant
structural differences - that are too subtle to be quantified and/or perceived
by a skilled human observer - can be elucidated between the positive
control data (wild-type vs. drug-treated) as well as between the
test data (wild-type vs. gene knockout).
Specifications
and Features
- Automated
3D image analysis tools in the MATLAB environment.
- Multiple
features automatically computed in batch mode.
- Statistical
comparison of data sets.
- Tools
will enable large-scale studies of protein deficiencies on the
mitotic spindle.
Fact
Sheet available here in PDF format.
Point
of Contact:
Jeffery R. Price, Ph.D.
Image Science & Machine Vision
Engineering Science and Technology Division
Oak Ridge National Laboratory
P.O. Box 2008, MS-6010
Oak Ridge, Tennessee 37831-6010
Office: (865) 574-5743
E-mail: pricejr@ornl.gov
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