Microstructured Block Copolymer Surfaces for Control of Microbe Adhesion and Aggregation

Hansen, Ryan R.; Shubert, Katherine R.; Morrell‐Falvey, Jennifer L.; Lokitz, Bradley S.; Doktycz, Mitchel J.; Retterer, Scott T.

doi:10.3390/bios4010063

Cited by 10 publications

(7 citation statements)

References 21 publications

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“…We showed patterning as close as 120 lm, but this could be decreased using optimized patterns and microfluidics, automated alignment of the fluidic device to the stencil layer (manual alignment was used here), or chemical surface modification. We did observe edge effects as has been reported previously, 32 and presumably this could be harnessed to improve pattern resolution. The resolution presented here is better than contemporary methods of pin-deposition or ink-jet printing.…”

Section: Discussionsupporting

confidence: 82%

Microstencils to generate defined, multi-species patterns of bacteria

et al. 2015

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Microbial communities are complex heterogeneous systems that are influenced by physical and chemical interactions with their environment, host, and community members. Techniques that facilitate the quantitative evaluation of how microscale organization influences the morphogenesis of multispecies communities could provide valuable insights into the dynamic behavior and organization of natural communities, the design of synthetic environments for multispecies culture, and the engineering of artificial consortia. In this work, we demonstrate a method for patterning microbes into simple arrangements that allow the quantitative measurement of growth dynamics as a function of their proximity to one another. The method combines parylene-based liftoff techniques with microfluidic delivery to simultaneously pattern multiple bacterial species with high viability using low-cost, customizable methods. Quantitative measurements of bacterial growth for two competing isolates demonstrate that spatial coordination can play a critical role in multispecies growth and structure. V C 2015 AIP Publishing LLC. [http://dx

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Section: Discussionsupporting

confidence: 82%

Microstencils to generate defined, multi-species patterns of bacteria

et al. 2015

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“…Control of the physical and chemical features of the microwell interface can be used to bias the bacterial populations isolated during the seeding step. It is well known that the attachment of bacterial cells to a solid interface depends on topological and chemical surface features, and numerous reports have used micro and nanofabrication strategies to control these properties, either to promote or inhibit bacteria attachment and interactions [ 34 – 36 ]. Here, substrates containing arrays of wells with diameters ranging from 5 to 1000 μm were used to investigate the effect of diameter on the population distribution of seeded Escherichia coli cells expressing green fluorescent protein (GFP).…”

Section: Resultsmentioning

confidence: 99%

Stochastic Assembly of Bacteria in Microwell Arrays Reveals the Importance of Confinement in Community Development

Hansen¹,

Timm²,

Timm³

et al. 2016

PLoS ONE

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The structure and function of microbial communities is deeply influenced by the physical and chemical architecture of the local microenvironment and the abundance of its community members. The complexity of this natural parameter space has made characterization of the key drivers of community development difficult. In order to facilitate these characterizations, we have developed a microwell platform designed to screen microbial growth and interactions across a wide variety of physical and initial conditions. Assembly of microbial communities into microwells was achieved using a novel biofabrication method that exploits well feature sizes for control of innoculum levels. Wells with incrementally smaller size features created populations with increasingly larger variations in inoculum levels. This allowed for reproducible growth measurement in large (20 μm diameter) wells, and screening for favorable growth conditions in small (5, 10 μm diameter) wells. We demonstrate the utility of this approach for screening and discovery using 5 μm wells to assemble P. aeruginosa colonies across a broad distribution of innoculum levels, and identify those conditions that promote the highest probability of survivial and growth under spatial confinement. Multi-member community assembly was also characterized to demonstrate the broad potential of this platform for studying the role of member abundance on microbial competition, mutualism and community succession.

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“…These properties are difficult to replicate in a bulk liquid culture or on an agar plate. The availability of microfluidic, micropatterning, and nanofabrication techniques that allow for the replication of key physical and chemical features of natural environments has, however, enabled many researchers to build bacterial communities to study their interactions 12,13,14 and to develop synthetic environments that mimic natural conditions 4,15,16,17,18,19,20 .…”

Section: Introductionmentioning

confidence: 99%

Assembly and Tracking of Microbial Community Development within a Microwell Array Platform

Timm

Halsted

Wilmoth

et al. 2017

JoVE

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The development of microbial communities depends on a combination of complex deterministic and stochastic factors that can dramatically alter the spatial distribution and activities of community members. We have developed a microwell array platform that can be used to rapidly assemble and track thousands of bacterial communities in parallel. This protocol highlights the utility of the platform and describes its use for optically monitoring the development of simple, two-member communities within an ensemble of arrays within the platform. This demonstration uses two mutants of Pseudomonas aeruginosa, part of a series of mutants developed to study Type VI secretion pathogenicity. Chromosomal inserts of either mCherry or GFP genes facilitate the constitutive expression of fluorescent proteins with distinct emission wavelengths that can be used to monitor community member abundance and location within each microwell. This protocol describes a detailed method for assembling mixtures of bacteria into the wells of the array and using time-lapse fluorescence imaging and quantitative image analysis to measure the relative growth of each member population over time. The seeding and assembly of the microwell platform, the imaging procedures necessary for the quantitative analysis of microbial communities within the array, and the methods that can be used to reveal interactions between microbial species area all discussed.

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Microstructured Block Copolymer Surfaces for Control of Microbe Adhesion and Aggregation

Cited by 10 publications

References 21 publications

Microstencils to generate defined, multi-species patterns of bacteria

Microstencils to generate defined, multi-species patterns of bacteria

Stochastic Assembly of Bacteria in Microwell Arrays Reveals the Importance of Confinement in Community Development

Assembly and Tracking of Microbial Community Development within a Microwell Array Platform

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