Microfluidic devices for studying growth and detachment of Staphylococcus epidermidis biofilms

Lee, Joung-Hyun; Kaplan, Jeffrey B.; Lee, Woo Y.

doi:10.1007/s10544-007-9157-0

Cited by 90 publications

(74 citation statements)

References 41 publications

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“…A device was developed to study the biofilm morphology forming under shear by imposing different flow velocities [21]. An integrated microfluidic chip has also been used for monitoring cell culture, including the monitoring of culture density over time and the probing cellular functions at a single-cell level [22].…”

Section: Introductionmentioning

confidence: 99%

Trends of Automation of Integrated Microfluidics

Lam¹

2013

Adv Robot Autom

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Section: Introductionmentioning

confidence: 99%

Trends of Automation of Integrated Microfluidics

Lam¹

2013

Adv Robot Autom

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“…In an effort to combine small volumes, high-throughput ability, and shear force in an open system, researchers have proposed the use of microfluidic models to study biofilms (4)(5)(6)(7). Some models have yet to demonstrate their high-throughput potential (5,7), while others require the system to be fabricated in the laboratory, which may not be feasible for all biofilm researchers (4-7).…”

mentioning

confidence: 99%

“…Some models have yet to demonstrate their high-throughput potential (5,7), while others require the system to be fabricated in the laboratory, which may not be feasible for all biofilm researchers (4-7). The platform proposed by Benoit et al (4) has high-throughput capability and uses a specialized instrument, the BioFlux device, that is commercially available.…”

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confidence: 99%

High-Throughput Microfluidic Method To Study Biofilm Formation and Host-Pathogen Interactions in Pathogenic Escherichia coli

Tremblay

Vogeleer

Jacques

et al. 2015

Appl Environ Microbiol

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Biofilm formation and host-pathogen interactions are frequently studied using multiwell plates; however, these closed systems lack shear force, which is present at several sites in the host, such as the intestinal and urinary tracts. Recently, microfluidic systems that incorporate shear force and very small volumes have been developed to provide cell biology models that resemble in vivo conditions. Therefore, the objective of this study was to determine if the BioFlux 200 microfluidic system could be used to study host-pathogen interactions and biofilm formation by pathogenic Escherichia coli. Strains of various pathotypes were selected to establish the growth conditions for the formation of biofilms in the BioFlux 200 system on abiotic (glass) or biotic (eukaryotic-cell) surfaces. Biofilm formation on glass was observed for the majority of strains when they were grown in M9 medium at 30°C but not in RPMI medium at 37°C. In contrast, HRT-18 cell monolayers enhanced binding and, in most cases, biofilm formation by pathogenic E. coli in RPMI medium at 37°C. As a proof of principle, the biofilm-forming ability of a diffusely adherent E. coli mutant strain lacking AIDA-I, a known mediator of attachment, was assessed in our models. In contrast to the parental strain, which formed a strong biofilm, the mutant formed a thin biofilm on glass or isolated clusters on HRT-18 monolayers. In conclusion, we describe a microfluidic method for high-throughput screening that could be used to identify novel factors involved in E. coli biofilm formation and host-pathogen interactions under shear force. Biofilms are defined as bacterial communities encased in a selfproduced polymeric matrix that is attached to a surface (1). The ability to form a biofilm is virtually a universal trait of bacteria and other microorganisms. Growth as a biofilm offers protection against hostile environments, the immune response, and bactericidal concentrations of antibiotics or disinfectants (1). Biofilms have frequently been studied using the 96-microtiter plate model because it is a platform that requires small volumes and is suited for high-throughput screening (1). The major flaws of the microtiter model are that it is a closed system and does not incorporate shear force. It is generally accepted that biofilms will develop in the presence of shear force in the environment (1). The MBEC assay, formerly known as the Calgary Biofilm Device, was developed to incorporate shear force into high-throughput screens (2); however, this model is a closed system. In a closed system, dispersing signals and metabolic waste accumulate, and nutrients become depleted. These events are not always favorable for biofilm studies and may lead to the rapid dispersal of the biofilm before it is quantified.Open systems, which have both a continuous flow of fresh medium and shear force, have been developed and are frequently used in the laboratory (1). These include the drip-flow reactor, flow cells, perfused biofilm fermenters, the CDC biofilm reactor, the rotating-disc re...

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“…Other microfluidic devices have recently been used for biofilm formation (14,19,21,23), but none of them has been used for HTP screening. The BioFlux device permits rapid measurement of the fluorescence of flow biofilms with a plate reader, which permits initial HTP screening of the viability of such biofilms.…”

mentioning

confidence: 99%

New Device for High-Throughput Viability Screening of Flow Biofilms

Benoit

Conant

Ionescu-Zanetti

et al. 2010

Appl Environ Microbiol

154

136

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Control of biofilms requires rapid methods to identify compounds effective against them and to isolate resistance-compromised mutants for identifying genes involved in enhanced biofilm resistance. While rapid screening methods for microtiter plate well ("static") biofilms are available, there are no methods for such screening of continuous flow biofilms ("flow biofilms"). Since the latter biofilms more closely approximate natural biofilms, development of a high-throughput (HTP) method for screening them is desirable. We describe here a new method using a device comprised of microfluidic channels and a distributed pneumatic pump (BioFlux) that provides fluid flow to 96 individual biofilms. This device allows fine control of continuous or intermittent fluid flow over a broad range of flow rates, and the use of a standard well plate format provides compatibility with plate readers. We show that use of green fluorescent protein (GFP)-expressing bacteria, staining with propidium iodide, and measurement of fluorescence with a plate reader permit rapid and accurate determination of biofilm viability. The biofilm viability measured with the plate reader agreed with that determined using plate counts, as well as with the results of fluorescence microscope image analysis. Using BioFlux and the plate reader, we were able to rapidly screen the effects of several antimicrobials on the viability of Pseudomonas aeruginosa PAO1 flow biofilms.Bacterial biofilms are surface-attached communities that are encased in a polymeric matrix, which exhibit a high degree of resistance to antimicrobial agents and the host immune system (12, 16). This makes them medically important; diseases with a biofilm component are chronic and difficult to eradicate. Examples of such diseases are cystitis (1), endocarditis (31), cystic fibrosis (35), and middle-ear (17) and indwelling medical device-associated (20) infections. Biofilms also play important environmental roles in, for example, wastewater treatment (38), bioremediation (29, 30), biofouling (7), and biocorrosion (2). Better control of biofilms requires elucidation of the molecular basis of their superior resistance (by identifying resistance-compromised mutants) and identification of compounds with antibiofilm activity. While our understanding of these aspects of biofilms has increased (11,15,(25)(26)(27)36), further work, including development of accurate high-throughput (HTP) methods for screening biofilm viability, is needed.Two major biofilm models are studied in the laboratory, biofilms grown without a continuous flow of fresh medium and biofilms grown with a continuous flow of fresh medium; examples of these two models are microtiter well biofilms and flow cell biofilms, respectively. Methods have been developed for HTP screening of the viability of static biofilms (6,28,32,33), but there are no methods for HTP screening of flow biofilms. The latter biofilms are typically grown in flow cells, which have to be examined individually to determine viability and thus cannot be used for ra...

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Microfluidic devices for studying growth and detachment of Staphylococcus epidermidis biofilms

Cited by 90 publications

References 41 publications

Trends of Automation of Integrated Microfluidics

Trends of Automation of Integrated Microfluidics

High-Throughput Microfluidic Method To Study Biofilm Formation and Host-Pathogen Interactions in Pathogenic Escherichia coli

New Device for High-Throughput Viability Screening of Flow Biofilms

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