Abstract
The mitotic spindle is a subcellular protein structure that, facilitates chromosome segregation and is crucial to cell division. We describe an image processing approach to quantitatively characterize and compare mitotic spindles that have been imaged three dimensionally using confocal microscopy with fixed-cell preparations. The proposed approach is based on a set of features that are computed from each image stack representing a spindle. We compare several spindle datasets of varying biological (genotype) and/or environmental (drug treatment) conditions. The goal of this effort is to aid biologists in detecting differences between spindles that may not be apparent under subjective visual inspection, and furthermore, to eventually automate such analysis in high-throughput scenarios (thousands of images) where manual inspection would be unreasonable. Experimental results on positive- and negative-control data indicate that the proposed approach is indeed effective. Differences are detected when it is known they do exist (positive control) and no differences are detected when there are none (negative control). In two other experimental comparisons, results indicate structural spindle differences that biologists had not observed previously.
Original language | English |
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Article number | 044012 |
Journal | Journal of Biomedical Optics |
Volume | 10 |
Issue number | 4 |
DOIs | |
State | Published - Jul 2005 |
Funding
The authors would like to acknowledge Lea Harrington and Bryan Snow, who directed the generation of a VPARP-deficient mouse model, and Dr. Atsushi Hirao for providing p53 deficient MEFs. We would like to thank Mark Ungrin, Philip Bingham, and Ken Tobin for their thorough reviews of this manuscript. We thank Conly Rieder for thoughtful and constructive discussions. We finally thank the anonymous reviewers whose constructive critiques helped us to significantly improve this work. Research sponsored by the Laboratory Directed Research and Development Program of Oak Ridge National Laboratory (ORNL), managed by UT-Battelle, LLC for the U.S. Department of Energy under contract number DE-AC05-00OR22725.
Keywords
- Confocal microscopy
- Fluorescence microscopy
- Gene knockout
- Image analysis
- Mitotic spindle
- Subcellular imaging