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1.
Biophys J ; 121(11): 2046-2059, 2022 06 07.
Article in English | MEDLINE | ID: mdl-35526093

ABSTRACT

To swim up gradients of nutrients, E. coli senses nutrient concentrations within its periplasm. For small nutrient molecules, periplasmic concentrations typically match extracellular concentrations. However, this is not necessarily the case for saccharides, such as maltose, which are transported into the periplasm via a specific porin. Previous observations have shown that, under various conditions, E. coli limits maltoporin abundance so that, for extracellular micromolar concentrations of maltose, there are predicted to be only nanomolar concentrations of free maltose in the periplasm. Thus, in the micromolar regime, the total uptake of maltose from the external environment into the cytoplasm is limited not by the abundance of cytoplasmic transport proteins but by the abundance of maltoporins. Here, we present results from experiments and modeling suggesting that this porin-limited transport enables E. coli to sense micromolar gradients of maltose despite having a high-affinity ABC transport system that is saturated at these micromolar levels. We used microfluidic assays to study chemotaxis of E. coli in various gradients of maltose and methyl-aspartate and leveraged our experimental observations to develop a mechanistic transport-and-sensing chemotaxis model. Incorporating this model into agent-based simulations, we discover a trade-off between uptake and sensing: although high-affinity transport enables higher uptake rates at low nutrient concentrations, it severely limits the range of dynamic sensing. We thus propose that E. coli may limit periplasmic uptake to increase its chemotactic sensitivity, enabling it to use maltose as an environmental cue.


Subject(s)
Escherichia coli Proteins , Periplasmic Binding Proteins , Bacterial Proteins/metabolism , Chemotaxis , Escherichia coli/metabolism , Escherichia coli Proteins/metabolism , Maltose/metabolism , Maltose-Binding Proteins/metabolism , Periplasmic Binding Proteins/metabolism , Porins/metabolism
2.
Sci Rep ; 11(1): 19508, 2021 09 30.
Article in English | MEDLINE | ID: mdl-34593946

ABSTRACT

Herein, we demonstrate that the use of index-matching materials (IMMs) allows direct visualization of microbial cells maintained at a solid-liquid interface through confocal reflection microscopy (CRM). The refractive index mismatch induces a background reflection at the solid-liquid interface that dwarfs the reflection signals from the cells and results in low-contrast images. We found that the IMMs sufficiently suppressed the background reflection at the solid-liquid interface, facilitating the imaging of microbes at the solid surface using CRM. The use of IMMs allowed quantitative analysis of the morphology of the mesh-like structure of Pseudomonas aeruginosa biofilms formed under denitrifying conditions, which led us to propose a novel structural model of the highly porous biofilm structure. These results indicate that the use of CRM coupled with an IMM offers a unique and promising tool for probing the dynamics of biofilm formation, along with visualization of environmental organisms and newly isolated bacteria, for which transformation methods are difficult to establish.


Subject(s)
Bacteria/cytology , Biofilms , Microbiota , Microscopy, Confocal/methods , Surface Properties
3.
Phys Biol ; 18(5)2021 06 23.
Article in English | MEDLINE | ID: mdl-33462162

ABSTRACT

Bacterial biofilms are communities of bacteria that exist as aggregates that can adhere to surfaces or be free-standing. This complex, social mode of cellular organization is fundamental to the physiology of microbes and often exhibits surprising behavior. Bacterial biofilms are more than the sum of their parts: single-cell behavior has a complex relation to collective community behavior, in a manner perhaps cognate to the complex relation between atomic physics and condensed matter physics. Biofilm microbiology is a relatively young field by biology standards, but it has already attracted intense attention from physicists. Sometimes, this attention takes the form of seeing biofilms as inspiration for new physics. In this roadmap, we highlight the work of those who have taken the opposite strategy: we highlight the work of physicists and physical scientists who use physics to engage fundamental concepts in bacterial biofilm microbiology, including adhesion, sensing, motility, signaling, memory, energy flow, community formation and cooperativity. These contributions are juxtaposed with microbiologists who have made recent important discoveries on bacterial biofilms using state-of-the-art physical methods. The contributions to this roadmap exemplify how well physics and biology can be combined to achieve a new synthesis, rather than just a division of labor.


Subject(s)
Bacterial Adhesion/physiology , Bacterial Physiological Phenomena , Biofilms , Quorum Sensing/physiology , Biofilms/growth & development
4.
Proc Natl Acad Sci U S A ; 117(41): 25571-25579, 2020 10 13.
Article in English | MEDLINE | ID: mdl-32973087

ABSTRACT

Optimal foraging theory provides a framework to understand how organisms balance the benefits of harvesting resources within a patch with the sum of the metabolic, predation, and missed opportunity costs of foraging. Here, we show that, after accounting for the limited environmental information available to microorganisms, optimal foraging theory and, in particular, patch use theory also applies to the behavior of marine bacteria in particle seascapes. Combining modeling and experiments, we find that the marine bacterium Vibrio ordalii optimizes nutrient uptake by rapidly switching between attached and planktonic lifestyles, departing particles when their nutrient concentration is more than hundredfold higher than background. In accordance with predictions from patch use theory, single-cell tracking reveals that bacteria spend less time on nutrient-poor particles and on particles within environments that are rich or in which the travel time between particles is smaller, indicating that bacteria tune the nutrient concentration at detachment to increase their fitness. A mathematical model shows that the observed behavioral switching between exploitation and dispersal is consistent with foraging optimality under limited information, namely, the ability to assess the harvest rate of nutrients leaking from particles by molecular diffusion. This work demonstrates how fundamental principles in behavioral ecology traditionally applied to animals can hold right down to the scale of microorganisms and highlights the exquisite adaptations of marine bacterial foraging. The present study thus provides a blueprint for a mechanistic understanding of bacterial uptake of dissolved organic matter and bacterial production in the ocean-processes that are fundamental to the global carbon cycle.


Subject(s)
Appetitive Behavior/physiology , Models, Biological , Organic Chemicals/metabolism , Plankton/physiology , Vibrio/physiology , Cyclic GMP , Geologic Sediments , Particulate Matter
5.
J Vis Exp ; (159)2020 05 27.
Article in English | MEDLINE | ID: mdl-32538911

ABSTRACT

Described here is confocal reflection microscopy-assisted single-cell innate fluorescence analysis (CRIF), a minimally invasive method for reconstructing the innate cellular fluorescence signature from each individual live cell in a population distributed in a three-dimensional (3D) space. The innate fluorescence signature of a cell is a collection of fluorescence signals emitted by various biomolecules within the cell. Previous studies established that innate fluorescence signatures reflect various cellular properties and differences in physiological status and are a rich source of information for cell characterization and identification. Innate fluorescence signatures have been traditionally analyzed at the population level, necessitating a clonal culture, but not at the single-cell level. CRIF is particularly suitable for studies that require 3D resolution and/or selective extraction of fluorescence signals from individual cells. Because the fluorescence signature is an innate property of a cell, CRIF is also suitable for tag-free prediction of the type and/or physiological status of intact and single cells. This method may be a powerful tool for streamlined cell analysis, where the phenotype of each single cell in a heterogenous population can be directly assessed by its autofluorescence signature under a microscope without cell tagging.


Subject(s)
Microscopy, Confocal , Single-Cell Analysis , Fluorescence , Microscopy, Confocal/methods , Pseudomonas putida , Saccharomyces cerevisiae , Single-Cell Analysis/methods
6.
J Vis Exp ; (155)2020 01 31.
Article in English | MEDLINE | ID: mdl-32065137

ABSTRACT

We demonstrate a method for the generation of controlled, dynamic chemical pulses-where localized chemoattractant becomes suddenly available at the microscale-to create micro-environments for microbial chemotaxis experiments. To create chemical pulses, we developed a system to introduce amino acid sources near-instantaneously by photolysis of caged amino acids within a polydimethylsiloxane (PDMS) microfluidic chamber containing a bacterial suspension. We applied this method to the chemotactic bacterium, Vibrio ordalii, which can actively climb these dynamic chemical gradients while being tracked by video microscopy. Amino acids, rendered biologically inert ('caged') by chemical modification with a photoremovable protecting group, are uniformly present in the suspension but not available for consumption until their sudden release, which occurs at user-defined points in time and space by means of a near-UV-A focused LED beam. The number of molecules released in the pulse can be determined by a calibration relationship between exposure time and uncaging fraction, where the absorption spectrum after photolysis is characterized by using UV-Vis spectroscopy. A nanoporous polycarbonate (PCTE) membrane can be integrated into the microfluidic device to allow the continuous removal by flow of the uncaged compounds and the spent media. A strong, irreversible bond between the PCTE membrane and the PDMS microfluidic structure is achieved by coating the membrane with a solution of 3-aminopropyltriethoxysilane (APTES) followed by plasma activation of the surfaces to be bonded. A computer-controlled system can generate user-defined sequences of pulses at different locations and with different intensities, so as to create resource landscapes with prescribed spatial and temporal variability. In each chemical landscape, the dynamics of bacterial movement at the individual scale and their accumulation at the population level can be obtained, thereby allowing the quantification of chemotactic performance and its effects on bacterial aggregations in ecologically relevant environments.


Subject(s)
Lab-On-A-Chip Devices/standards , Microfluidics/instrumentation , Humans
7.
Appl Environ Microbiol ; 85(18)2019 09 15.
Article in English | MEDLINE | ID: mdl-31324624

ABSTRACT

Here we analyzed the innate fluorescence signature of the single microbial cell, within both clonal and mixed populations of microorganisms. We found that even very similarly shaped cells differ noticeably in their autofluorescence features and that the innate fluorescence signatures change dynamically with growth phases. We demonstrated that machine learning models can be trained with a data set of single-cell innate fluorescence signatures to annotate cells according to their phenotypes and physiological status, for example, distinguishing a wild-type Aspergillus nidulans cell from its nitrogen metabolism mutant counterpart and log-phase cells from stationary-phase cells of Pseudomonas putida We developed a minimally invasive method (confocal reflection microscopy-assisted single-cell innate fluorescence [CRIF] analysis) to optically extract and catalog the innate cellular fluorescence signatures of each of the individual live microbial cells in a three-dimensional space. This technique represents a step forward from traditional techniques which analyze the innate fluorescence signatures at the population level and necessitate a clonal culture. Since the fluorescence signature is an innate property of a cell, our technique allows the prediction of the types or physiological status of intact and tag-free single cells, within a cell population distributed in a three-dimensional space. Our study presents a blueprint for a streamlined cell analysis where one can directly assess the potential phenotype of each single cell in a heterogenous population by its autofluorescence signature under a microscope, without cell tagging.IMPORTANCE A cell's innate fluorescence signature is an assemblage of fluorescence signals emitted by diverse biomolecules within a cell. It is known that the innate fluoresce signature reflects various cellular properties and physiological statuses; thus, they can serve as a rich source of information in cell characterization as well as cell identification. However, conventional techniques focus on the analysis of the innate fluorescence signatures at the population level but not at the single-cell level and thus necessitate a clonal culture. In the present study, we developed a technique to analyze the innate fluorescence signature of a single microbial cell. Using this novel method, we found that even very similarly shaped cells differ noticeably in their autofluorescence features, and the innate fluorescence signature changes dynamically with growth phases. We also demonstrated that the different cell types can be classified accurately within a mixed population under a microscope at the resolution of a single cell, depending solely on the innate fluorescence signature information. We suggest that single-cell autofluoresce signature analysis is a promising tool to directly assess the taxonomic or physiological heterogeneity within a microbial population, without cell tagging.


Subject(s)
Aspergillus fumigatus/physiology , Fluorescence , Machine Learning , Microscopy, Confocal/methods , Pseudomonas putida/physiology , Single-Cell Analysis/methods
8.
Proc Natl Acad Sci U S A ; 116(22): 10792-10797, 2019 05 28.
Article in English | MEDLINE | ID: mdl-31097577

ABSTRACT

Ephemeral aggregations of bacteria are ubiquitous in the environment, where they serve as hotbeds of metabolic activity, nutrient cycling, and horizontal gene transfer. In many cases, these regions of high bacterial concentration are thought to form when motile cells use chemotaxis to navigate to chemical hotspots. However, what governs the dynamics of bacterial aggregations is unclear. Here, we use an experimental platform to create realistic submillimeter-scale nutrient pulses with controlled nutrient concentrations. By combining experiments, mathematical theory, and agent-based simulations, we show that individual Vibrio ordalii bacteria begin chemotaxis toward hotspots of dissolved organic matter (DOM) when the magnitude of the chemical gradient rises sufficiently far above the sensory noise that is generated by stochastic encounters with chemoattractant molecules. Each DOM hotspot is surrounded by a dynamic ring of chemotaxing cells, which congregate in regions of high DOM concentration before dispersing as DOM diffuses and gradients become too noisy for cells to respond to. We demonstrate that V. ordalii operates close to the theoretical limits on chemotactic precision. Numerical simulations of chemotactic bacteria, in which molecule counting noise is explicitly taken into account, point at a tradeoff between nutrient acquisition and the cost of chemotactic precision. More generally, our results illustrate how limits on sensory precision can be used to understand the location, spatial extent, and lifespan of bacterial behavioral responses in ecologically relevant environments.


Subject(s)
Bacteria , Chemotaxis/physiology , Models, Biological , Bacteria/drug effects , Bacteria/metabolism , Chemotactic Factors/pharmacology , Computer Simulation , Environment , Signal-To-Noise Ratio , Vibrio/drug effects , Vibrio/physiology
9.
ISME J ; 13(3): 563-575, 2019 03.
Article in English | MEDLINE | ID: mdl-30446738

ABSTRACT

Aquatic environments harbor a great diversity of microorganisms, which interact with the same patchy, particulate, or diffuse resources by means of a broad array of physiological and behavioral adaptations, resulting in substantially different life histories and ecological success. To date, efforts to uncover and understand this diversity have not been matched by equivalent efforts to identify unifying frameworks that can provide a degree of generality and thus serve as a stepping stone to scale up microscale dynamics to predict their ecosystem-level consequences. In particular, evaluating the ecological consequences of different resource landscapes and of different microbial adaptations has remained a major challenge in aquatic microbial ecology. Here, inspired by Ramon Margalef's mandala for phytoplankton, we propose a foraging mandala for microorganisms in aquatic environments, which accounts for both the local environment and individual adaptations. This biophysical framework distills resource acquisition into two fundamental parameters: the search time for a new resource and the growth return obtained from encounter with a resource. We illustrate the foraging mandala by considering a broad range of microbial adaptations and environmental characteristics. The broad applicability of the foraging mandala suggests that it could be a useful framework to compare disparate microbial strategies in aquatic environments and to reduce the vast complexity of microbe-environment interactions into a minimal number of fundamental parameters.


Subject(s)
Bacteria , Hydrobiology , Microbial Interactions , Phytoplankton/physiology , Adaptation, Physiological , Bacterial Physiological Phenomena , Ecosystem
10.
Sci Rep ; 6: 33115, 2016 09 21.
Article in English | MEDLINE | ID: mdl-27650454

ABSTRACT

Quorum sensing (QS) is a population-density dependent chemical process that enables bacteria to communicate based on the production, secretion and sensing of small inducer molecules. While recombinant constructs have been widely used to decipher the molecular details of QS, how those findings translate to natural QS systems has remained an open question. Here, we compare the activation of natural and synthetic Pseudomonas aeruginosa LasI/R QS systems in bacteria exposed to quiescent conditions and controlled flows. Quantification of QS-dependent GFP expression in suspended cultures and in surface-attached microcolonies revealed that QS onset in both systems was similar under quiescent conditions but markedly differed under flow. Moderate flow (Pe > 25) was sufficient to suppress LasI/R QS recombinantly expressed in Escherichia coli, whereas only high flow (Pe > 102) suppressed QS in wild-type P. aeruginosa. We suggest that this difference stems from the differential production of extracellular matrix and that the matrix confers resilience against moderate flow to QS in wild-type organisms. These results suggest that the expression of a biofilm matrix extends the environmental conditions under which QS-based cell-cell communication is effective and that findings from synthetic QS circuits cannot be directly translated to natural systems.


Subject(s)
Escherichia coli/physiology , Pseudomonas aeruginosa/metabolism , Quorum Sensing , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Escherichia coli/genetics , Gene Expression Regulation, Bacterial , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Recombinant Proteins/metabolism
11.
J Bacteriol ; 198(19): 2589-95, 2016 10 01.
Article in English | MEDLINE | ID: mdl-27274032

ABSTRACT

The advent of microscale technologies, such as microfluidics, has revolutionized many areas of biology yet has only recently begun to impact the field of bacterial biofilms. By enabling accurate control and manipulation of physical and chemical conditions, these new microscale approaches afford the ability to combine important features of natural and artificial microbial habitats, such as fluid flow and ephemeral nutrient sources, with an unprecedented level of flexibility and quantification. Here, we review selected case studies to exemplify this potential, discuss limitations, and suggest that this approach opens new vistas into biofilm research over traditional setups, allowing us to expand our understanding of the formation and consequences of biofilms in a broad range of environments and applications.


Subject(s)
Biofilms/growth & development , Microfluidic Analytical Techniques , Bacterial Adhesion , Environment , Time Factors
12.
Biosci Biotechnol Biochem ; 80(1): 7-12, 2016.
Article in English | MEDLINE | ID: mdl-26103134

ABSTRACT

Cells respond to the environment and alter gene expression. Recent studies have revealed the social aspects of bacterial life, such as biofilm formation. Biofilm formation is largely affected by the environment, and the mechanisms by which the gene expression of individual cells affects biofilm development have attracted interest. Environmental factors determine the cell's decision to form or leave a biofilm. In addition, the biofilm structure largely depends on the environment, implying that biofilms are shaped to adapt to local conditions. Second messengers such as cAMP and c-di-GMP are key factors that link environmental factors with gene regulation. Cell-to-cell communication is also an important factor in shaping the biofilm. In this short review, we will introduce the basics of biofilm formation and further discuss environmental factors that shape biofilm formation. Finally, the state-of-the-art tools that allow us investigate biofilms under various conditions are discussed.


Subject(s)
Bacterial Proteins/genetics , Biofilms/growth & development , Gene Expression Regulation, Bacterial , Gene-Environment Interaction , Pseudomonas aeruginosa/genetics , Second Messenger Systems/genetics , Bacillus subtilis/genetics , Bacillus subtilis/metabolism , Bacillus subtilis/ultrastructure , Bacterial Adhesion , Bacterial Proteins/metabolism , Clostridium perfringens/genetics , Clostridium perfringens/metabolism , Clostridium perfringens/ultrastructure , Cyclic AMP/metabolism , Cyclic GMP/analogs & derivatives , Cyclic GMP/metabolism , Microscopy, Electron, Scanning , Pseudomonas aeruginosa/metabolism , Pseudomonas aeruginosa/ultrastructure , Quorum Sensing/genetics , Species Specificity , Vibrio cholerae/genetics , Vibrio cholerae/metabolism , Vibrio cholerae/ultrastructure
13.
Biosci Biotechnol Biochem ; 78(1): 178-81, 2014.
Article in English | MEDLINE | ID: mdl-25036502

ABSTRACT

Biofilms are communities of surface-attached microbial cells that resist environmental stresses. In this study, we found that low concentrations of ethanol increase biofilm formation in Pseudomonas aeruginosa PAO1 but not in a mutant of it lacking both Psl and Pel exopolysaccharides. Low concentrations of ethanol also increased pellicle formation at the air-liquid interface.


Subject(s)
Biofilms/drug effects , Biofilms/growth & development , Ethanol/pharmacology , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/physiology , Dose-Response Relationship, Drug , Polysaccharides, Bacterial/metabolism , Pseudomonas aeruginosa/genetics , Pseudomonas aeruginosa/metabolism
14.
Proc Natl Acad Sci U S A ; 111(15): 5622-7, 2014 Apr 15.
Article in English | MEDLINE | ID: mdl-24706766

ABSTRACT

Although competition-dispersal tradeoffs are commonly invoked to explain species coexistence for animals and plants in spatially structured environments, such mechanisms for coexistence remain unknown for microorganisms. Here we show that two recently speciated marine bacterioplankton populations pursue different behavioral strategies to exploit nutrient particles in adaptation to the landscape of ephemeral nutrient patches characteristic of ocean water. These differences are mediated primarily by differential colonization of and dispersal among particles. Whereas one population is specialized to colonize particles by attaching and growing biofilms, the other is specialized to disperse among particles by rapidly detecting and swimming toward new particles, implying that it can better exploit short-lived patches. Because the two populations are very similar in their genomic composition, metabolic abilities, chemotactic sensitivity, and swimming speed, this fine-scale behavioral adaptation may have been responsible for the onset of the ecological differentiation between them. These results demonstrate that the principles of spatial ecology, traditionally applied at macroscales, can be extended to the ocean's microscale to understand how the rich spatiotemporal structure of the resource landscape contributes to the fine-scale ecological differentiation and species coexistence among marine bacteria.


Subject(s)
Bacterial Physiological Phenomena , Biofilms/growth & development , Chemotaxis/physiology , Demography , Genetic Speciation , Plankton/physiology , Chitin , Microfluidics , Microscopy, Confocal , Microscopy, Electron, Transmission , Microscopy, Fluorescence , Models, Biological , Oceans and Seas , Plankton/ultrastructure
15.
Microbiol Immunol ; 57(8): 589-93, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23647374

ABSTRACT

Biofilms, such as dental plaque, are aggregates of microorganisms attached to a surface. Thus, visualization of biofilms together with their attached substrata is important in order to understand details of the interaction between them. However, so far there is limited availability of such techniques. Here, non-invasive visualization of biofilm formation with its attached substratum by applying the previously reported technique of continuous-optimizing confocal reflection microscopy (COCRM) is reported. The process of development of oral biofilm together with its substratum was sequentially visualized with COCRM. This study describes a convenient method for visualizing biofilm and its attached surface.


Subject(s)
Biofilms , Dental Plaque/microbiology , Microscopy, Confocal/methods , Mouth/microbiology , Streptococcus mutans/physiology , Bacterial Adhesion , Humans , Streptococcus mutans/chemistry
16.
Microbes Environ ; 28(1): 13-24, 2013.
Article in English | MEDLINE | ID: mdl-23363620

ABSTRACT

Microbes interact with each other in multicellular communities and this interaction enables certain microorganisms to survive in various environments. Pseudomonas aeruginosa is a highly adaptable bacterium that ubiquitously inhabits diverse environments including soil, marine habitats, plants and animals. Behind this adaptivity, P. aeruginosa has abilities not only to outcompete others but also to communicate with each other to develop a multispecies community. In this review, we focus on how P. aeruginosa interacts with other microorganisms. P. aeruginosa secretes antimicrobial chemicals to compete and signal molecules to cooperate with other organisms. In other cases, it directly conveys antimicrobial enzymes to other bacteria using the Type VI secretion system (T6SS) or membrane vesicles (MVs). Quorum sensing is a central regulatory system used to exert their ability including antimicrobial effects and cooperation with other microbes. At least three quorum sensing systems are found in P. aeruginosa, Las, Rhl and Pseudomonas quinolone signal (PQS) systems. These quorum-sensing systems control the synthesis of extracellular antimicrobial chemicals as well as interaction with other organisms via T6SS or MVs. In addition, we explain the potential of microbial interaction analysis using several micro devices, which would bring fresh sensitivity to the study of interspecies interaction between P. aeruginosa and other organisms.


Subject(s)
Microbial Interactions , Pseudomonas aeruginosa/physiology , Anti-Infective Agents/metabolism , Gene Expression Regulation, Bacterial , Pseudomonas aeruginosa/genetics , Pseudomonas aeruginosa/metabolism , Quorum Sensing , Signal Transduction , Species Specificity , Transport Vesicles/physiology
17.
Analyst ; 138(4): 1000-3, 2013 Feb 21.
Article in English | MEDLINE | ID: mdl-23289096

ABSTRACT

A microfluidic device was developed for rapid determination of the minimum inhibitory concentration (MIC) of antibiotics against bacteria. A small volume of sample solution was introduced into multiple chambers simultaneously, and the growth of bacteria was quantified using a noninvasive three-dimensional (3D) visualization technique.


Subject(s)
Anti-Bacterial Agents/pharmacology , Escherichia coli/drug effects , Microfluidic Analytical Techniques/methods , Anti-Bacterial Agents/chemistry , Escherichia coli/physiology , Microbial Sensitivity Tests/instrumentation , Microbial Sensitivity Tests/methods , Microfluidic Analytical Techniques/instrumentation , Time Factors
18.
Appl Environ Microbiol ; 77(12): 4253-5, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21515728

ABSTRACT

Continuous monitoring of ammonia removal by microbial complexes and observation of their morphology were carried out using a microdevice. Consumption of NH(4)(+) ions by active sludge could clearly be recorded over 48 h. Aggregation of the sludge could be observed in parallel by using confocal reflection microscopy.


Subject(s)
Ammonia/metabolism , Microbiological Techniques/methods , Sewage/microbiology , Microscopy, Confocal/methods
19.
J Biosci Bioeng ; 110(3): 377-80, 2010 Sep.
Article in English | MEDLINE | ID: mdl-20547370

ABSTRACT

The feasibility of a method to monitor biofilm development non-destructively in a microfluidic device was addressed. Here, we report that biofilm growth could be non-destructively monitored by an image analysis technique based on modification of confocal reflection microscopy.


Subject(s)
Bacterial Load/methods , Biofilms/growth & development , Image Interpretation, Computer-Assisted/methods , Microfluidic Analytical Techniques/methods , Microscopy, Confocal/methods , Streptococcus mutans/cytology , Streptococcus mutans/growth & development , Bacterial Load/instrumentation , Bioreactors , Cell Culture Techniques , Microfluidic Analytical Techniques/instrumentation
20.
J Biosci Bioeng ; 110(1): 130-3, 2010 Jul.
Article in English | MEDLINE | ID: mdl-20541131

ABSTRACT

The feasibility of a method to nondestructively measure planktonic bacterial growth in a microfluidic device was addressed. Here, we report that the growth of Pseudomonas aeruginosa in a microfluidic device could be measured by a three-dimensional image analysis technique based on confocal reflection microscopy in a time-course.


Subject(s)
Bacteriological Techniques/instrumentation , Bacteriological Techniques/methods , Microfluidic Analytical Techniques , Microscopy, Confocal , Pseudomonas aeruginosa/growth & development , DNA , Imaging, Three-Dimensional/methods
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