Vibrio cholerae biofilm growth program and architecture revealed by single-cell live imaging. Author Jing Yan, Andrew Sharo, Howard Stone, Ned Wingreen, Bonnie Bassler Publication Year 2016 Type Journal Article Abstract Biofilms are surface-associated bacterial communities that are crucial in nature and during infection. Despite extensive work to identify biofilm components and to discover how they are regulated, little is known about biofilm structure at the level of individual cells. Here, we use state-of-the-art microscopy techniques to enable live single-cell resolution imaging of a Vibrio cholerae biofilm as it develops from one single founder cell to a mature biofilm of 10,000 cells, and to discover the forces underpinning the architectural evolution. Mutagenesis, matrix labeling, and simulations demonstrate that surface adhesion-mediated compression causes V. cholerae biofilms to transition from a 2D branched morphology to a dense, ordered 3D cluster. We discover that directional proliferation of rod-shaped bacteria plays a dominant role in shaping the biofilm architecture in V. cholerae biofilms, and this growth pattern is controlled by a single gene, rbmA Competition analyses reveal that the dense growth mode has the advantage of providing the biofilm with superior mechanical properties. Our single-cell technology can broadly link genes to biofilm fine structure and provides a route to assessing cell-to-cell heterogeneity in response to external stimuli. Keywords Bacterial Adhesion, Biofilms, Vibrio cholerae, Bacterial Proteins, Humans, Cell Proliferation, Single-Cell Analysis Journal Proc Natl Acad Sci U S A Volume 113 Issue 36 Pages E5337-43 Date Published 2016 Sep 06 ISSN Number 1091-6490 DOI 10.1073/pnas.1611494113 Alternate Journal Proc Natl Acad Sci U S A PMCID PMC5018804 PMID 27555592 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML