Abstract
During the life of a cell, numerous essential cellular processes must be coordinated both spatially and temporally, from DNA replication and chromosome segregation to gene expression and cytokinesis. In order to analyze these inherently dynamic and cell-cycle-dependent processes, it is essential to observe the dynamic localization of the cellular machinery throughout the entire cell cycle. Although some coarse features of cell-cycle dynamics can be captured in snapshot imaging, where cellular size or morphology can be used as a proxy for cell-cycle phase, the inherently stochastic nature of ultrastructures in the cell makes the direct visualization of subcellular dynamics an essential tool to differentiate between structural differences that are the result of biologically relevant dynamics versus cell-to-cell variation. With these goals in mind, we have developed a unique high-throughput imaging approach, and have recently applied this to characterize the cell-cycle localization of nearly every protein in the bacterial cell (Kuwada in Mol Microbiol, 95(1), 64–79, 2015). This approach combines large-format sample preparation with automated image capture, processing, and analysis to quantitatively characterize proteome localization of tens of thousands of complete cell cycles.
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Communicated by M. Kupiec.
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Kuwada, N.J., Traxler, B. & Wiggins, P.A. High-throughput cell-cycle imaging opens new doors for discovery. Curr Genet 61, 513–516 (2015). https://doi.org/10.1007/s00294-015-0493-y
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DOI: https://doi.org/10.1007/s00294-015-0493-y