Thrust and Power Output of the Bacterial Flagellar Motor: A Micromagnetic Tweezers Approach.

One of the most common swimming strategies employed by microorganisms is based on the use of rotating helical filaments, called flagella, that are powered by molecular motors. Determining the physical properties of this propulsive system is crucial to understanding the behavior of these organisms. Furthermore, the ability to dynamically monitor the activity of the flagellar motor is a valuable indicator of the overall energetics of the cell. In this work, inherently magnetic bacteria confined in micromagnetic CoFe traps are used to directly and noninvasively determine the flagellar thrust force and swimming speed of motile cells. The technique permits determination of the ratio of propulsive force/swimming speed (the hydrodynamic resistance) and the power output of the flagellar motor for individual cells over extended time periods. Cells subjected to ultraviolet radiation are observed to experience exponential decays in power output as a function of exposure time. By noninvasively measuring thrust, velocity, and power output over time at a single-cell level, this technique can serve as the foundation for fundamental studies of bacterial hydrodynamics and also provides a novel, to our knowledge, tether-free probe of single-cell energetics over time.

Magnetic tweezers-based 3D microchannel electroporation for high-throughput gene transfection in living cells.

A novel high-throughput magnetic tweezers-based 3D microchannel electroporation system capable of transfecting 40 000 cells/cm(2) on a single chip for gene therapy, regenerative medicine, and intracellular detection of target mRNA for screening cellular heterogeneity is reported. A single cell or an ordered array of individual cells are remotely guided by programmable magnetic fields to poration sites with high (>90%) cell alignment efficiency to enable various transfection reagents to be delivered simultaneously into the cells. The present technique, in contrast to the conventional vacuum-based approach, is significantly gentler on the cellular membrane yielding >90% cell viability and, moreover, allows transfected cells to be transported for further analysis. Illustrating the versatility of the system, the GATA2 molecular beacon is delivered into leukemia cells to detect the regulation level of the GATA2 gene that is associated with the initiation of leukemia. The uniform delivery and a sharp contrast of fluorescence intensity between GATA2 positive and negative cells demonstrate key aspects of the platform for gene transfer, screening and detection of targeted intracellular markers in living cells.

A MagDot-Nanoconveyor Assay Detects and Isolates Molecular Biomarkers.

The ability to quickly analyze, separate, and manipulate multiple types of biomarkers from small sample volumes is a significant step toward personalized medicine.

Real-time magnetic actuation of DNA nanodevices via modular integration with stiff micro-levers.

DNA nanotechnology has enabled complex nanodevices, but the ability to directly manipulate systems with fast response times remains a key challenge. Current methods of actuation are relatively slow and only direct devices into one or two target configurations. Here we report an approach to control DNA origami assemblies via externally applied magnetic fields using a low-cost platform that enables actuation into many distinct configurations with sub-second response times. The nanodevices in these assemblies are manipulated via mechanically stiff micron-scale lever arms, which rigidly couple movement of a micron size magnetic bead to reconfiguration of the nanodevice while also enabling direct visualization of the conformation. We demonstrate control of three assemblies-a rod, rotor, and hinge-at frequencies up to several Hz and the ability to actuate into many conformations. This level of spatiotemporal control over DNA devices can serve as a foundation for real-time manipulation of molecular and atomic systems.

Dynamic trajectory analysis of superparamagnetic beads driven by on-chip micromagnets.

We investigate the non-linear dynamics of superparamagnetic beads moving around the periphery of patterned magnetic disks in the presence of an in-plane rotating magnetic field. Three different dynamical regimes are observed in experiments, including (1) phase-locked motion at low driving frequencies, (2) phase-slipping motion above the first critical frequency fc1, and (3) phase-insulated motion above the second critical frequency fc2. Experiments with Janus particles were used to confirm that the beads move by sliding rather than rolling. The rest of the experiments were conducted on spherical, isotropic magnetic beads, in which automated particle position tracking algorithms were used to analyze the bead dynamics. Experimental results in the phase-locked and phase-slipping regimes correlate well with numerical simulations. Additional assumptions are required to predict the onset of the phase-insulated regime, in which the beads are trapped in closed orbits; however, the origin of the phase-insulated state appears to result from local magnetization defects. These results indicate that these three dynamical states are universal properties of bead motion in non-uniform oscillators.

On-chip magnetic separation and encapsulation of cells in droplets.

Single cell study is gaining importance because of the cell-to-cell variation that exists within cell population, even after significant initial sorting. Analysis of such variation at the gene expression level could impact single cell functional genomics, cancer, stem-cell research, and drug screening. The on-chip monitoring of individual cells in an isolated environment would prevent cross-contamination, provide high recovery yield, and enable study of biological traits at a single cell level. These advantages of on-chip biological experiments is a significant improvement for a myriad of cell analyses methods, compared to conventional methods, which require bulk samples and provide only averaged information on cell structure and function. We report on a device that integrates a mobile magnetic trap array with microfluidic technology to provide the possibility of separation of immunomagnetically labeled cells and their encapsulation with reagents into picoliter droplets for single cell analysis. The simultaneous reagent delivery and compartmentalization of the cells immediately following sorting are all performed seamlessly within the same chip. These steps offer unique advantages such as the ability to capture cell traits as originated from its native environment, reduced chance of contamination, minimal use of the reagents, and tunable encapsulation characteristics independent of the input flow. Preliminary assay on cell viability demonstrates the potential for the device to be integrated with other up- or downstream on-chip modules to become a powerful single-cell analysis tool.

Ultra-localized single cell electroporation using silicon nanowires.

Analysis of cell-to-cell variation can further the understanding of intracellular processes and the role of individual cell function within a larger cell population. The ability to precisely lyse single cells can be used to release cellular components to resolve cellular heterogeneity that might be obscured when whole populations are examined. We report a method to position and lyse individual cells on silicon nanowire and nanoribbon biological field effect transistors. In this study, HT-29 cancer cells were positioned on top of transistors by manipulating magnetic beads using external magnetic fields. Ultra-rapid cell lysis was subsequently performed by applying 600-900 mV(pp) at 10 MHz for as little as 2 ms across the transistor channel and the bulk substrate. We show that the fringing electric field at the device surface disrupts the cell membrane, leading to lysis from irreversible electroporation. This methodology allows rapid and simple single cell lysis and analysis with potential applications in medical diagnostics, proteome analysis and developmental biology studies.

Light-scattering study of the normal human eye lens: elastic properties and age dependence.

The human ocular lens is a tissue capable of changing its shape to dynamically adjust the optical power of the eye, a function known as accommodation, which gradually declines with age. This capability is the response of the lens tissue to external forces, which, in turn, is modulated by the biomechanical characteristics of lens tissues. In order to investigate the contributions of lens sclerosis to loss of accommodation, we report on in vitro confocal Brillouin light scattering studies of human ocular lenses spanning over a 30-70 year age range. Using this nondestructive measurement method, we determined that the longitudinal bulk modulus (average ± SD) of the lens nucleus (2.79 ± 0.14 GPa) was consistently greater than the bulk modulus of the lens cortex (2.36 ± 0.09 GPa). Moreover, our results showed that these differences were not age dependent over the 40 year age range that we evaluated using healthy lens tissues. Our results are consistent with the hypothesis that an age-dependent change in the bulk modulus of lens tissues does not fully account for the natural decline of accommodation.