title: High-throughput cell-cycle imaging opens new doors for discovery.
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.
authors.affiliation: Department of Physics, University of Washington, Seattle, WA, 98195, USA. nateman@uw.edu.|Department of Bioengineering, University of Washington, Seattle, WA, 98195, USA. nateman@uw.edu.
authors.forename: Nathan J
authors.initials: NJ
authors.lastname: Kuwada
authors.affiliation: Department of Microbiology, University of Washington, Seattle, WA, 98195, USA.
authors.forename: Beth
authors.initials: B
authors.lastname: Traxler
authors.affiliation: Department of Physics, University of Washington, Seattle, WA, 98195, USA.|Department of Bioengineering, University of Washington, Seattle, WA, 98195, USA.|Department of Microbiology, University of Washington, Seattle, WA, 98195, USA.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Wiggins
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D016935:Caulobacter crescentus / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure*; D002453:Cell Cycle / Q000235:genetics; D004261:DNA Replication; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure*; D015964:Gene Expression Regulation, Bacterial; D057166:High-Throughput Screening Assays; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation / Q000379:methods; D057054:Molecular Imaging / Q000295:instrumentation / Q000379:methods*; D020543:Proteome / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods; D059008:Time-Lapse Imaging / Q000295:instrumentation / Q000379:methods
chemical_list: D001426:Bacterial Proteins; D020543:Proteome; D011993:Recombinant Fusion Proteins
keywords: Bacterial protein dynamics; High-throughput imaging; Quantitative cell biology