Flagellated bacterial motility in polymer solutions Martinez, Vincent A.; Schwarz-Linek, Jana; Reufer, Mathias ...
Proceedings of the National Academy of Sciences - PNAS,
12/2014, Letnik:
111, Številka:
50
Journal Article
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Significance The way microorganisms swim in concentrated polymer solutions has important biomedical implications, i.e., how pathogens invade the mucosal lining of mammal guts. Physicists are also ...fascinated by self-propulsion in such complex non-Newtonian fluids. The current standard model of how bacteria propelled by rotary helical flagella swim through concentrated polymer solutions postulates bacteria-sized pores, allowing them relative easy passage. Our experiments using high-throughput methods overturn this standard model. Instead, we show that the peculiarities of flagellated bacteria locomotion in concentrated polymer solutions are due to the fast-rotating flagellum, giving rise to a lower local viscosity in its vicinity. The bacterial flagellum is therefore a nano-rheometer for probing flows at the molecular level.
It is widely believed that the swimming speed, v , of many flagellated bacteria is a nonmonotonic function of the concentration, c , of high-molecular-weight linear polymers in aqueous solution, showing peaked Formula curves. Pores in the polymer solution were suggested as the explanation. Quantifying this picture led to a theory that predicted peaked Formula curves. Using high-throughput methods for characterizing motility, we measured v and the angular frequency of cell body rotation, Ω, of motile Escherichia coli as a function of polymer concentration in polyvinylpyrrolidone (PVP) and Ficoll solutions of different molecular weights. We find that nonmonotonic Formula curves are typically due to low-molecular-weight impurities. After purification by dialysis, the measured Formula and Formula relations for all but the highest-molecular-weight PVP can be described in detail by Newtonian hydrodynamics. There is clear evidence for non-Newtonian effects in the highest-molecular-weight PVP solution. Calculations suggest that this is due to the fast-rotating flagella seeing a lower viscosity than the cell body, so that flagella can be seen as nano-rheometers for probing the non-Newtonian behavior of high polymer solutions on a molecular scale.
Photogravitactic Microswimmers Singh, Dhruv P.; Uspal, William E.; Popescu, Mihail N. ...
Advanced functional materials,
June 20, 2018, Letnik:
28, Številka:
25
Journal Article
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Phototactic microorganisms are commonly observed to respond to natural sunlight by swimming upward against gravity. This study demonstrates that synthetic photochemically active microswimmers can ...also swim against gravity. The particles initially sediment and, when illuminated at low light intensities exhibit wall‐bound states of motion near the bottom surface. Upon increasing the intensity of light, the artificial swimmers lift off from the wall and swim against gravity and away from the light source. This motion in the bulk has been further confirmed using holographic microscopy. A theoretical model is presented within the framework of self‐diffusiophoresis, which allows to unequivocally identify the photochemical activity and the phototactic response as key mechanisms in the observed phenomenology. Since the lift‐off threshold intensity depends on the particle size, it can be exploited to selectively address particles with the same density from a polydisperse mixture of active particles and move them in or out of the boundary region. This study provides a simple design strategy to fabricate artificial microswimmers whose two‐ or three‐dimensional swimming behavior can be controlled with light.
Photogravitactic swimming of artificial microswimmers. Photochemically active Janus colloids, initially sedimented to a bottom surface, swim along the surface when illuminated with UV light at low intensity. Upon increasing the light intensity above certain threshold value, they lift‐off and move against gravity and away from the light source, a behavior bearing similarities with that of microorganisms such as phyto and zooplankton.
We present a fast, high-throughput method for characterizing the motility of microorganisms in three dimensions based on standard imaging microscopy. Instead of tracking individual cells, we analyze ...the spatiotemporal fluctuations of the intensity in the sample from time-lapse images and obtain the intermediate scattering function of the system. We demonstrate our method on two different types of microorganisms: the bacterium Escherichia coli (both smooth swimming and wild type) and the biflagellate alga Chlamydomonas reinhardtii. We validate the methodology using computer simulations and particle tracking. From the intermediate scattering function, we are able to extract the swimming speed distribution, fraction of motile cells, and diffusivity for E. coli, and the swimming speed distribution, and amplitude and frequency of the oscillatory dynamics for C. reinhardtii. In both cases, the motility parameters were averaged over ∼104 cells and obtained in a few minutes.
Sperm require a sense of direction to locate the egg for fertilization. They follow gradients of chemical and physical cues provided by the egg or the oviduct. However, the principles underlying ...three-dimensional (3D) navigation in chemical landscapes are unknown. Here using holographic microscopy and optochemical techniques, we track sea urchin sperm navigating in 3D chemoattractant gradients. Sperm sense gradients on two timescales, which produces two different steering responses. A periodic component, resulting from the helical swimming, gradually aligns the helix towards the gradient. When incremental path corrections fail and sperm get off course, a sharp turning manoeuvre puts sperm back on track. Turning results from an 'off' Ca(2+) response signifying a chemoattractant stimulation decrease and, thereby, a drop in cyclic GMP concentration and membrane voltage. These findings highlight the computational sophistication by which sperm sample gradients for deterministic klinotaxis. We provide a conceptual and technical framework for studying microswimmers in 3D chemical landscapes.
Bacterial flagella are helical proteinaceous fibers, composed of the protein flagellin, that confer motility to many bacterial species. The genomes of about half of all flagellated species include ...more than one flagellin gene, for reasons mostly unknown. Here we show that two flagellins (FlaA and FlaB) are spatially arranged in the polar flagellum of Shewanella putrefaciens, with FlaA being more abundant close to the motor and FlaB in the remainder of the flagellar filament. Observations of swimming trajectories and numerical simulations demonstrate that this segmentation improves motility in a range of environmental conditions, compared to mutants with single-flagellin filaments. In particular, it facilitates screw-like motility, which enhances cellular spreading through obstructed environments. Similar mechanisms may apply to other bacterial species and may explain the maintenance of multiple flagellins to form the flagellar filament.
Axonemes form the core of eukaryotic flagella and cilia, performing tasks ranging from transporting fluid in developing embryos to the propulsion of sperm. Despite their abundance across the ...eukaryotic domain, the mechanisms that regulate the beating action of axonemes remain unknown. The flagellar waveforms are 3D in general, but current understanding of how axoneme components interact stems from 2D data; comprehensive measurements of flagellar shape are beyond conventional microscopy. Moreover, current flagellar model systems (e.g., sea urchin, human sperm) contain accessory structures that impose mechanical constraints on movement, obscuring the “native” axoneme behavior. We address both problems by developing a high-speed holographic imaging scheme and applying it to the (male) microgametes of malaria (Plasmodium) parasites. These isolated flagella are a unique, mathematically tractable model system for the physics of microswimmers. We reveal the 3D flagellar waveforms of these microorganisms and map the differential shear between microtubules in their axonemes. Furthermore, we overturn claims that chirality in the structure of the axoneme governs the beat pattern Hirokawa N, et al. (2009) Ann Rev Fluid Mech 41:53–72, because microgametes display a left- or right-handed character on alternate beats. This breaks the link between structural chirality in the axoneme and larger scale symmetry breaking (e.g., in developing embryos), leading us to conclude that accessory structures play a critical role in shaping the flagellar beat.
The three-dimensional swimming tracks of motile microorganisms can be used to identify their species, which holds promise for the rapid identification of bacterial pathogens. The tracks also provide ...detailed information on the cells' responses to external stimuli such as chemical gradients and physical objects. Digital holographic microscopy (DHM) is a well-established, but computationally intensive method for obtaining three-dimensional cell tracks from video microscopy data. We demonstrate that a common neural network (NN) accelerates the analysis of holographic data by an order of magnitude, enabling its use on single-board computers and in real time. We establish a heuristic relationship between the distance of a cell from the focal plane and the size of the bounding box assigned to it by the NN, allowing us to rapidly localise cells in three dimensions as they swim. This technique opens the possibility of providing real-time feedback in experiments, for example by monitoring and adapting the supply of nutrients to a microbial bioreactor in response to changes in the swimming phenotype of microbes, or for rapid identification of bacterial pathogens in drinking water or clinical samples.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
We introduce active, probe-based microrheological techniques for measuring the flow and deformation of complex fluids. These techniques are ideal for mechanical characterization either when little ...sample is available, or when samples show significant spatial heterogeneity. We review recent results, paying particular attention to comparing and contrasting rheological parameters obtained from micro- and macro-rheological techniques.
Archaea have evolved to survive in some of the most extreme environments on earth. Life in extreme, nutrient-poor conditions gives the opportunity to probe fundamental energy limitations on movement ...and response to stimuli, two essential markers of living systems. Here we use three-dimensional holographic microscopy and computer simulations to reveal that halophilic archaea achieve chemotaxis with power requirements one hundred-fold lower than common eubacterial model systems. Their swimming direction is stabilised by their flagella (archaella), enhancing directional persistence in a manner similar to that displayed by eubacteria, albeit with a different motility apparatus. Our experiments and simulations reveal that the cells are capable of slow but deterministic chemotaxis up a chemical gradient, in a biased random walk at the thermodynamic limit.
Many motile microorganisms are able to detect chemical gradients in their surroundings to bias their motion toward more favorable conditions. In this study, we observe the swimming patterns of ...Caulobacter crescentus, a uniflagellated bacterium, in a linear oxygen gradient produced by a three-channel microfluidic device. Using low-magnification dark-field microscopy, individual cells are tracked over a large field of view and their positions within the oxygen gradient are recorded over time. Motor switching events are identified so that swimming trajectories are deconstructed into a series of forward and backward swimming runs. Using these data, we show that C. crescentus displays aerotactic behavior by extending the average duration of forward swimming runs while moving up an oxygen gradient, resulting in directed motility toward oxygen sources. Additionally, the motor switching response is sensitive both to the steepness of the gradient experienced and to background oxygen levels, exhibiting a logarithmic response.