is supported by a NIH Cellular, Biochemical, and Molecular Sciences (CMB) training grant (T32GM007276). are supported by the Allen Discovery Center at Stanford on Systems Modeling of Infection ( ). is supported by a Burroughs Wellcome Fund Career Award at the Scientific Interface (CASI) ( ), and a Beckman Young Investigator Award ( ). This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the paper and its Supporting Information files.įunding: Y.L., M.K., and B.W. Received: ApAccepted: SeptemPublished: October 17, 2019Ĭopyright: © 2019 Lim et al. PLoS Biol 17(10):Īcademic Editor: Raghuveer Parthasarathy, The University of Oregon, UNITED STATES (2019) Mechanically resolved imaging of bacteria using expansion microscopy. Focusing on host–microbe interactions that are difficult to quantify through fluorescence alone, we demonstrate the ability of μExM to distinguish species through an in vitro defined community of human gut commensals and in vivo imaging of a model gut microbiota, and to sensitively detect cell-envelope damage caused by antibiotics or previously unrecognized cell-to-cell phenotypic heterogeneity among pathogenic bacteria as they infect macrophages.Ĭitation: Lim Y, Shiver AL, Khariton M, Lane KM, Ng KM, Bray SR, et al. We use this phenomenon as a quantitative and sensitive phenotypic imaging contrast orthogonal to spectral separation to resolve bacterial cells of different species or in distinct physiological states. We find that expansion patterns depend on the structural and mechanical properties of the cell wall, which vary across species and conditions. While prior studies have relied on techniques involving spectral labeling, we have developed an expansion microscopy method (μExM) in which bacterial cells are physically expanded prior to imaging. Imaging dense and diverse microbial communities has broad applications in basic microbiology and medicine, but remains a grand challenge due to the fact that many species adopt similar morphologies.
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