Intra-Species Bacterial Quorum Sensing Studied at Single Cell Level in a Double Droplet Trapping System

Bai, Yunpeng; Patil, Santoshkumar N.; Bowden, Steven D.; Poulter, Simon; Pan, Jie; Salmond, George P. C.; Welch, Martin; Huck, Wilhelm T. S.; Abell, Chris

doi:10.3390/ijms140510570

Cited by 24 publications

(17 citation statements)

References 22 publications

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“…Extending these findings to predict the activity within more complex multispecies microbial ecosystems will require a combination of predictive theoretical models and perhaps new experimental tools to quantify interactions. With the rapid development of nanofabrication technology, microfluidic devices are widely used for single-cell analysis [ 41 , 42 ], allowing us to study multispecies interactions at the microsystem level such as suggested in Fig 4 and elsewhere [ 43 , 44 ]. We implemented a model in Fig 4 to explore how the species evenness and the interaction network set the overall ecosystem outputs.…”

Section: Discussionmentioning

confidence: 99%

The Contribution of High-Order Metabolic Interactions to the Global Activity of a Four-Species Microbial Community

Guo

Boedicker

2016

PLoS Comput Biol

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The activity of a biological community is the outcome of complex processes involving interactions between community members. It is often unclear how to accurately incorporate these interactions into predictive models. Previous work has shown a range of positive and negative metabolic pairwise interactions between species. Here we examine the ability of a modified general Lotka-Volterra model with cell-cell interaction coefficients to predict the overall metabolic rate of a well-mixed microbial community comprised of four heterotrophic natural isolates, experimentally quantifying the strengths of two, three, and four-species interactions. Within this community, interactions between any pair of microbial species were positive, while higher-order interactions, between 3 or more microbial species, slightly modulated community metabolism. For this simple community, the metabolic rate of can be well predicted only with taking into account pairwise interactions. Simulations using the experimentally determined interaction parameters revealed that spatial heterogeneity in the distribution of cells increased the importance of multispecies interactions in dictating function at both the local and global scales.

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Section: Discussionmentioning

confidence: 99%

The Contribution of High-Order Metabolic Interactions to the Global Activity of a Four-Species Microbial Community

Guo

Boedicker

2016

PLoS Comput Biol

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“…However, upon their adjustment to prolonged mammalian assays, they require multilayer fabrication, either involving high-pressure cell loading or not allowing for different substrates to be used (26,27). Several single-cell platforms that encapsulate cells in stationary droplet arrays are available (5,(28)(29)(30)(31)(32)(33)(34)(35); however, the ability to culture adherent cells for long periods remains difficult (36).…”

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confidence: 99%

Stationary nanoliter droplet array with a substrate of choice for single adherent/nonadherent cell incubation and analysis

Shemesh

Ben-Arye

Avesar³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Microfluidic water-in-oil droplets that serve as separate, chemically isolated compartments can be applied for single-cell analysis; however, to investigate encapsulated cells effectively over prolonged time periods, an array of droplets must remain stationary on a versatile substrate for optimal cell compatibility. We present here a platform of unique geometry and substrate versatility that generates a stationary nanodroplet array by using wells branching off a main microfluidic channel. These droplets are confined by multiple sides of a nanowell and are in direct contact with a biocompatible substrate of choice. The device is operated by a unique and reversed loading procedure that eliminates the need for fine pressure control or external tubing. Fluorocarbon oil isolates the droplets and provides soluble oxygen for the cells. By using this approach, the metabolic activity of single adherent cells was monitored continuously over time, and the concentration of viable pathogens in blood-derived samples was determined directly by measuring the number of colony-formed droplets. The method is simple to operate, requires a few microliters of reagent volume, is portable, is reusable, and allows for cell retrieval. This technology may be particularly useful for multiplexed assays for which prolonged and simultaneous visual inspection of many isolated single adherent or nonadherent cells is required.single cell | nanoliter array | diagnostics C ommon single-cell analysis methods, such as flow cytometry and mass cytometry (1), offer high throughput and accurate single-cell marker quantification, yet they lack the ability to monitor large numbers of single cells continuously and simultaneously in performance-based assays (2, 3). Conventional microscopy may be used for these assays; however, in the case of single cells, they cannot analyze extracellular events, such as secretion. To achieve this, cells must be isolated in compartments that can sustain cell viability and growth while permitting conventional optical analysis over many hours to days. Dropletbased microfluidics, which enables single-cell encapsulation in nano-and subnanoliter droplets by surrounding microscopic aqueous medium with an immiscible carrier fluid (4-8), recently gained interest with the appearance of digital PCR (9-11). Much of the work thus far has been directed toward improving droplet manipulation capabilities (12-16). With these methods, droplets are mobile, and thus cytometry is performed under flow conditions (17), making continuous monitoring of single cells difficult. Continuous monitoring may be achieved by using stationary indexed droplets, but many current droplet immobilization techniques are limited by pressure coupling between droplet generation and capture events, as well as the requirement to adjust droplet volume to nanowell size (6,18,19). The vast majority of methods used to generate water-in-oil droplets begin by priming a continuous oil phase in a microfluidic channel followed by an injection of a dispersed (aqueous) medium (2...

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“…2013). In E. coli, quorum sensing was investigated at the single-cell scale using a double droplet-based microfluidic system enabling to encapsulate two individual cells separately into two adjacent droplets which can communicate by means of diffusion of signal molecules across the droplets (Bai et al, 2013). Using strains carrying a GFP construction under the control of the quorum sensing receptor protein LasR, the activation of quorum sensing was analyzed via the monitoring of GFP expression in an individual cell in a droplet in response to autoinducers produced by the cell in the adjacent droplet.…”

Section: Resident Biofilms As Hot Spots For Pathogen Persistencementioning

confidence: 99%

Fluorescence-based tools for single-cell approaches in food microbiology

Bridier¹,

Hammes

Canette

et al. 2015

International Journal of Food Microbiology

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Intra-Species Bacterial Quorum Sensing Studied at Single Cell Level in a Double Droplet Trapping System

Cited by 24 publications

References 22 publications

The Contribution of High-Order Metabolic Interactions to the Global Activity of a Four-Species Microbial Community

The Contribution of High-Order Metabolic Interactions to the Global Activity of a Four-Species Microbial Community

Stationary nanoliter droplet array with a substrate of choice for single adherent/nonadherent cell incubation and analysis

Fluorescence-based tools for single-cell approaches in food microbiology

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