Book description: At long last, here is the thoroughly revised and updated, and long-anticipated, third edition of the hugely successful Art of Electronics. Widely accepted as the best single authoritative text on electronic circuit design, it will be an indispensable reference and the gold standard for anyone in the field.
Weakly-scattering objects, such as small colloidal particles and most biological cells, are frequently encountered in microscopy. Indeed, a range of techniques have been developed to better visualize these phase objects; phase contrast and DIC are among the most popular methods for enhancing contrast. However, recording position and shape in the out-of-imaging-plane direction remains challenging. This report introduces a simple experimental method to accurately determine the location and geometry of objects in three dimensions, using digital inline holographic microscopy (DIHM). Broadly speaking, the accessible sample volume is defined by the camera sensor size in the lateral direction, and the illumination coherence in the axial direction. Typical sample volumes range from 200 µm x 200 µm x 200 µm using LED illumination, to 5 mm x 5 mm x 5 mm or larger using laser illumination. This illumination light is configured so that plane waves are incident on the sample. Objects in the sample volume then scatter light, which interferes with the unscattered light to form interference patterns perpendicular to the illumination direction. This image (the hologram) contains the depth information required for three-dimensional reconstruction, and can be captured on a standard imaging device such as a CMOS or CCD camera. The Rayleigh-Sommerfeld back propagation method is employed to numerically refocus microscope images, and a simple imaging heuristic based on the Gouy phase anomaly is used to identify scattering objects within the reconstructed volume. This simple but robust method results in an unambiguous, model-free measurement of the location and shape of objects in microscopic samples.
Swimming bacteria explore their environment by performing a random walk, which is biased in response to, for example, chemical stimuli, resulting in a collective drift of bacterial populations towards ‘a better life’. This phenomenon, called chemotaxis, is one of the best known forms of collective behaviour in bacteria, crucial for bacterial survival and virulence. Both single-cell and macroscopic assays have investigated bacterial behaviours. However, theories that relate the two scales have previously been difficult to test directly. We present an image analysis method, inspired by light scattering, which measures the average collective motion of thousands of bacteria simultaneously. Using this method, a time-varying collective drift as small as 50 nm s−1 can be measured. The method, validated using simulations, was applied to chemotactic Escherichia colibacteria in linear gradients of the attractant α-methylaspartate. This enabled us to test a coarse-grained minimal model of chemotaxis. Our results clearly map the onset of receptor methylation, and the transition from linear to logarithmic sensing in the bacterial response to an external chemoeffector. Our method is broadly applicable to problems involving the measurement of collective drift with high time resolution, such as cell migration and fluid flows measurements, and enables fast screening of tactic behaviours.
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 v(c) curves. Pores in the polymer solution were suggested as the explanation. Quantifying this picture led to a theory that predicted peaked v(c) 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 v(c) curves are typically due to low-molecular-weight impurities. After purification by dialysis, the measured v(c) and Ω(c) 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.
We demonstrate an innovative multifunctional artificial material that combines exotic metamaterial properties and the environmentally responsive nature of phase change media. The tunable metamaterial is designed with the aid of two interwoven coordinate-transformation equations and implemented with a network of thin film resistors and vanadium dioxide (VO2). The strong temperature dependence of VO2 electrical conductivity results in a relevant modification of the resistor network behavior, and we provide experimental evidence for a reconfigurable metamaterial electric circuit (MMEC) that not only mimics a continuous medium but is also capable of responding to thermal stimulation through dynamic variation of its spatial anisotropy. Upon external temperature change the overall effective functionality of the material switches between a "truncated-cloak" and "concentrator" for electric currents. Possible applications may include adaptive matching resistor networks, multifunctional electronic devices, and equivalent artificial materials in the magnetic domain. Additionally, the proposed technology could also be relevant for thermal management of integrated circuits
Combination therapy has become one of the leading approaches for treating complex diseases because it coadministers clinically proven drugs to concurrently target multiple signaling pathways of diseased cells. Identification of synergic drug combinations at their respective effective doses without unwanted accumulative side effects is the key to success for such therapy. In this work, we demonstrate the feasibility of the vortex-assisted microfluidic electroporation system for direct drug cocktail analyses where drug substances were individually delivered into cytosols in a sequential and dosage-controlled manner. Through quantitative analyses, the synergic combinational dosage ratios of the chemotherapeutic drug and the anticancer flavonoid were identified. When integrated with high-throughput label-free rare cell purification techniques, the presented system has the potential for development of personalized medicines as the system would be capable of comprehensively assessing drug combinations directly on patients’ cellular samples.
Electroporation has received increasing attention in the past years, because it is a very powerful technique for physically introducing non-permeant exogenous molecular probes into cells. This work reports a microfluidic electroporation platform capable of performing multiple molecule delivery to mammalian cells with precise and molecular-dependent parameter control. The system’s ability to isolate cells with uniform size distribution allows for less variation in electroporation efficiency per given electric field strength; hence enhanced sample viability. Moreover, its process visualization feature allows for observation of the fluorescent molecular uptake process in real-time, which permits prompt molecular delivery parameter adjustments in situ for efficiency enhancement. To show the vast capabilities of the reported platform, macromolecules with different sizes and electrical charges (e.g., Dextran with MW of 3,000 and 70,000 Da) were delivered to metastatic breast cancer cells with high delivery efficiencies (>70%) for all tested molecules. The developed platform has proven its potential for use in the expansion of research fields where on-chip electroporation techniques can be beneficial.
Schonbrun E, Malka R, Caprio GD, Schaak D, Higgins JM.
Digital cameras would be colorblind if they did not have pixelated color filters integrated into their image sensors. Integration of conventional fixed filters, however, comes at the expense of an inability to modify the camera’s spectral properties. Instead, we demonstrate a micropolarizer-based camera that can reconfigure its spectral response. Color is encoded into a linear polarization state by a chiral dispersive element and then read out in a single exposure. The polarization encoded color camera is capable of capturing three-color images at wavelengths spanning the visible to the near infrared.
Spatial tailoring of the material constitutive properties is a well-known strategy to mold the local flow of given observables in different physical domains. Coordinate-transformation-based methods (e.g., trans-formation optics) offer a powerful and systematic approach to design anisotropic, spatially inhomogeneous artificial materials (metamaterials) capable of precisely manipulating wave-based (electromagnetic, acoustic, elastic) as well as diffusion-based (heat) phenomena in a desired fashion. However, as versatile as these approaches have been, most designs have thus far been limited to serving single-target functionalities in a given physical domain. Here, we present a step towards a “transformation multiphysics” framework that allows independent and simultaneous manipulation of multiple physical phenomena. As a proof of principle of this new scheme, we design and synthesize (in terms of realistic material constituents) a metamaterial shell that simultaneously behaves as a thermal concentrator and an electrical “invisibility cloak.” Our numerical results open up intriguing possibilities in the largely unexplored phase space of multifunctional metadevices, with a wide variety of potential applications to electrical, magnetic, acoustic, and thermal scenarios.
. Comptes Rendus de l’Academie des Sciences 2014;15(10):898-906.Abstract
The Sagnac effect has played an instrumental role for the fundamental studies of relativity, and various devices that utilize this effect have been applied to many disciplines ranging from inertial navigation to geodesy and to seismology. In this context we present an overview of recent developments related to superfluid helium quantum interference devices. With the discovery of superfluid Josephson phenomena in He4, the device technology has been rapidly developing in the past 10 years. We discuss the underlying working principles of these interference devices and their applications. We focus on their use as sensitive rotation sensors based on the Sagnac effect coupled with the existence of a macroscopic quantum phase via particle–wave duality.
Microbial communities abound with examples of complex social interactions that shape microbial ecosystems. One particularly striking example is microbial cooperation via the secretion of public goods. It has been suggested by theory, and recently demonstrated experimentally, that microbial population dynamics and the evolutionary dynamics of cooperative social genes take place with similar timescales, and are linked to each other via an eco-evolutionary feedback loop. We overview this recent evidence, and discuss the possibility that a third process may be also part of this loop: phenotypic dynamics. Complex social strategies may be implemented at the single-cell level by means of gene regulatory networks. Thus gene expression plasticity or stochastic gene expression, both of which may occur with a timescale of one to a few generations, can potentially lead to a three-way coupling between behavioral dynamics, population dynamics, and evolutionary dynamics
Ecosystems can undergo sudden shifts to undesirable states, but recent studies with simple single-species ecosystems have demonstrated that advance warning can be provided by the slowing down of population dynamics near a tipping point. However, it is unclear how this ‘critical slowing down’ will manifest in ecosystems with strong interactions between their components. Here we probe the dynamics of an experimental producer-freeloader ecosystem as it approaches a catastrophic collapse. Surprisingly, the producer population grows in size as the environment deteriorates, highlighting that population size can be a misleading measure of ecosystem stability. By analysing the oscillatory producer-freeloader dynamics for over 100 generations in multiple environmental conditions, we find that the collective ecosystem dynamics slow down as the tipping point is approached. Analysis of the coupled dynamics of interacting populations may therefore be necessary to provide advance warning of collapse in complex communities.
Artificial materials engineered to exhibit properties that do not typically exist in nature are often called metamaterials, and these unique materials have attracted a significant amount of scientific interest in recent years. Just as conventional materials owe their properties to the average response from an ensemble of atoms and molecules, in “artificial” materials, each structural unit plays the role of an atom. The material properties are controlled not only by what elements are used and their individual properties, but also by the way they are arranged collectively in lattice-like geometrical patterns with prescribed spatial variations. This immense flexibility is one of the main reasons why material engineering has become such an explosive catalyst in so many scientific disciplines. For example, metamaterials applied to the manipulation of electromagnetic waves have demonstrated some counterintuitive concepts such as a negative index of refraction, which allows light to bend at the interface of materials in a direction opposite to what you would observe from any ordinary materials. The concept of artificial material engineering has extended beyond conventional material science and it has now started to be applied to the manipulation of heat flow.
Application of force on biomolecules has been instrumental in understanding biofunctional behaviour from single molecules to complex collections of cells. Current approaches, for example, those based on atomic force microscopy or magnetic or optical tweezers, are powerful but limited in their applicability as integrated biosensors. Here we describe a new force-based biosensing technique based on the quartz crystal microbalance. By applying centrifugal forces to a sample, we show it is possible to repeatedly and non-destructively interrogate its mechanical properties in situ and in real time. We employ this platform for the studies of micron-sized particles, viscoelastic monolayers of DNA and particles tethered to the quartz crystal microbalance surface by DNA. Our results indicate that, for certain types of samples on quartz crystal balances, application of centrifugal force both enhances sensitivity and reveals additional mechanical and viscoelastic properties.
Imaging fluorescence in moving cells is fundamentally challenging because the exposure time is constrained by motion-blur, which limits the available signal. We report a method to image fluorescently labeled leukemia cells in fluid flow that has an effective exposure time of up to 50 times the motion-blur limit. Flowing cells are illuminated with a pseudo-random excitation pulse sequence, resulting in a motion-blur that can be computationally removed to produce near diffraction-limited images. This method enables observation of cellular organelles and their behavior in a fluid environment that resembles the vasculature.
In Drosophila, most individual olfactory receptor neurons (ORNs) project bilaterally to both sides of the brain. Having bilateral rather than unilateral projections may represent a useful redundancy. However, bilateral ORN projections to the brain should also compromise the ability to lateralize odours. Nevertheless, walking or flying Drosophila reportedly turn towards the antenna that is more strongly stimulated by odour. Here we show that each ORN spike releases approximately 40% more neurotransmitter from the axon branch ipsilateral to the soma than from the contralateral branch. As a result, when an odour activates the antennae asymmetrically, ipsilateral central neurons begin to spike a few milliseconds before contralateral neurons, and at a 30to50% higher rate than contralateral neurons. We show that a walking fly can detect a 5% asymmetry in total ORN input to its left and right antennal lobes, and can turn towards the odour in less time than it requires the fly to complete a stride. These results demonstrate that neurotransmitter release properties can be tuned independently at output synapses formed by a single axon onto two target cells with identical functions and morphologies. Our data also show that small differences in spike timing and spike rate can produce reliable differences in olfactory behaviour.
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.