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 ∼10(4) cells and obtained in a few minutes.
Utilizing a multilayered composite approach, we have designed and constructed a new class of artificial materials for thermal conduction. We show that an engineered material can be utilized to control the diffusive heat flow in ways inconceivable with naturally occurring materials. By shielding, concentrating, and inverting heat current, we experimentally demonstrate the unique potential and the utility of guiding heat flux.
Bacillus spores are highly resistant dormant cells formed in response to starvation. The spore is surrounded by a structurally complex protein shell, the coat, which protects the genetic material. In spite of its dormancy, once nutrient is available (or an appropriate physical stimulus is provided) the spore is able to resume metabolic activity and return to vegetative growth, a process requiring the coat to be shed. Spores dynamically expand and contract in response to humidity, demanding that the coat be flexible. Despite the coat's critical biological functions, essentially nothing is known about the design principles that allow the coat to be tough but also flexible and, when metabolic activity resumes, to be efficiently shed. Here, we investigated the hypothesis that these apparently incompatible characteristics derive from an adaptive mechanical response of the coat. We generated a mechanical model predicting the emergence and dynamics of the folding patterns uniformly seen in Bacillus spore coats. According to this model, spores carefully harness mechanical instabilities to fold into a wrinkled pattern during sporulation. Owing to the inherent nonlinearity in their formation, these wrinkles persist during dormancy and allow the spore to accommodate changes in volume without compromising structural and biochemical integrity. This characteristic of the spore and its coat may inspire design of adaptive materials.
We present an overview of recent developments related to superfluid helium quantum interference devices (SHeQUIDs). We discuss the physics of two reservoirs of superfluid helium coupled together and describe the quantum oscillations that result from varying the coupling strength. We explain the principles behind SHeQUIDs that can be built based on these oscillations and review some techniques and applications.
We present a simple and secure system for encrypting and decrypting information using DNA self-assembly. Binary data is encoded in the geometry of DNA nanostructures with two distinct conformations. Removing or leaving out a single component reduces these structures to an encrypted solution of ssDNA, whereas adding back this missing "decryption key" causes the spontaneous formation of the message through self-assembly, enabling rapid read out via gel electrophoresis. Applications include authentication, secure messaging, and barcoding.
Capture and isolation of flowing cells and particulates from body fluids has enormous implications in diagnosis, monitoring, and drug testing, yet monovalent adhesion molecules used for this purpose result in inefficient cell capture and difficulty in retrieving the captured cells. Inspired by marine creatures that present long tentacles containing multiple adhesive domains to effectively capture flowing food particulates, we developed a platform approach to capture and isolate cells using a 3D DNA network comprising repeating adhesive aptamer domains that extend over tens of micrometers into the solution. The DNA network was synthesized from a microfluidic surface by rolling circle amplification where critical parameters, including DNA graft density, length, and sequence, could readily be tailored. Using an aptamer that binds to protein tyrosine kinase-7 (PTK7) that is overexpressed on many human cancer cells, we demonstrate that the 3D DNA network significantly enhances the capture efficiency of lymphoblast CCRF-CEM cells over monovalent aptamers and antibodies, yet maintains a high purity of the captured cells. When incorporated in a herringbone microfluidic device, the 3D DNA network not only possessed significantly higher capture efficiency than monovalent aptamers and antibodies, but also outperformed previously reported cell-capture microfluidic devices at high flow rates. This work suggests that 3D DNA networks may have broad implications for detection and isolation of cells and other bioparticles.
Replicating bacterial chromosomes continuously demix from each other and segregate within a compact volume inside the cell called the nucleoid. Although many proteins involved in this process have been identified, the nature of the global forces that shape and segregate the chromosomes has remained unclear because of limited knowledge of the micromechanical properties of the chromosome. In this work, we demonstrate experimentally the fundamentally soft nature of the bacterial chromosome and the entropic forces that can compact it in a crowded intracellular environment. We developed a unique "micropiston" and measured the force-compression behavior of single Escherichia coli chromosomes in confinement. Our data show that forces on the order of 100 pN and free energies on the order of 10(5) k(B)T are sufficient to compress the chromosome to its in vivo size. For comparison, the pressure required to hold the chromosome at this size is a thousand-fold smaller than the surrounding turgor pressure inside the cell. Furthermore, by manipulation of molecular crowding conditions (entropic forces), we were able to observe in real time fast (approximately 10 s), abrupt, reversible, and repeatable compaction-decompaction cycles of individual chromosomes in confinement. In contrast, we observed much slower dissociation kinetics of a histone-like protein HU from the whole chromosome during its in vivo to in vitro transition. These results for the first time provide quantitative, experimental support for a physical model in which the bacterial chromosome behaves as a loaded entropic spring in vivo.
This study used a novel feedback approach to control a robotic foot using force and length signals transmitted from an isolated Xenopus laevis frog muscle. The foot's environment (inertial versus hydrodynamic), gearing (outlever/inlever) and size were changed to alter the muscle's load. Upon nerve stimulation (250 Hz, 80 ms train duration), variation in loading generated a range of muscle stress (19.8±5.3 to 66.0±22.5 kPa), work (1.89±0.67 to 6.87±2.96 J kg(-1) muscle) and power (12.4±7.5 to 64.8±28.3 W kg(-1) muscle; mean ± s.d., N=6 frogs). Inertial versus hydrodynamic loading dramatically shifted contractile dynamics. With the foot in water, the muscle generated ∼30% higher force, yet shortened slower, producing lower power than inertial loading. Power increased in air from 22.6±5.8 to 63.6±27.2 W kg(-1) muscle in response to doubling the gear ratio, but did not increase in water. Surprisingly, altering foot size diminished muscle performance in water, causing power to drop significantly from 41.6±11.1 to 25.1±8.0 W kg(-1) muscle as foot area was doubled. Thus, morphological modifications influenced muscle dynamics independently of neural control; however, changes in loading environment and gearing affected contractile output more strongly than changes in foot size. Confirming recent theory, these findings demonstrate how muscle contractile output can be modulated solely by altering the mechanical environment.
Materials that have subwavelength structure can add degrees of freedom to optical system design that are not possible with bulk materials. We demonstrate two lenses that are composed out of lithographically patterned arrays of elliptical cross-section silicon nanowires, which can dynamically reconfigure their imaging properties in response to the polarization of the illumination. In each element, two different focusing functions are polarization encoded into a single lens. The first nanowire lens has a different focal length for each linear polarization state, thereby realizing the front end of a nonmechanical zoom imaging system. The second nanowire lens has a different optical axis for each linear polarization state, demonstrating stereoscopic image capture from a single physical aperture.
Wrubel J, Gabrielse G, Klothammer WS, Larochelle P, McConnell R, Richerme P, Grzonka D, Oelert W, Sefzick T, Zielinski M, Borbely JS, George MC, Hessels EA, Storry CH, Weel M, Mullers A, Walz J, Speck A. Pumped helium system for cooling positron and electron traps to 1.2 K. In: Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment. 2011 p. 232–240.
Single-molecule techniques provide opportunities for molecularly precise imaging, manipulation, assembly and biophysical studies. Owing to the kinetics of bond rupture processes, rapid single-molecule measurements can reveal novel bond rupture mechanisms, probe single-molecule events with short lifetimes and enhance the interaction forces supplied by single molecules. Rapid measurements will also increase throughput necessary for technological use of single-molecule techniques. Here we report a nanomechanical sensor that allows single-molecule force spectroscopy on the previously unexplored microsecond timescale. We probed bond lifetimes around 5 μs and observed significant enhancements in molecular interaction forces. Our loading-rate-dependent measurements provide experimental evidence for an additional energy barrier in the biotin-streptavidin complex. We also demonstrate quantitative mapping of rapid single-molecule interactions with high spatial resolution. This nanomechanical interface may allow studies of molecular processes with short lifetimes and development of novel biological imaging, single-molecule manipulation and assembly technologies.
We report a new kind of experiment in which we take an array of nanoscale apertures that form a superfluid (4)He Josephson junction and apply quantum phase gradients directly along the array. We observe collective coherent behaviors from aperture elements, leading to quantum interference. Connections to superconducting and Bose-Einstein condensate Josephson junctions as well as phase coherence among the superfluid aperture array are discussed.
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.
We report the observation of superfluid quantum interference in a compact, large-area matter-wave interferometer consisting of a multiple-turn interfering path in reciprocal geometry. Utilizing the Sagnac effect from Earth's rotation in conjunction with a phase shifter made of superfluid heat current, we demonstrate that such a scheme can be extended for sensitive rotation sensing as well as for general interferometry.