Single particle tracking reveals biphasic transport during nanorod magnetophoresis through extracellular matrix

We quantify nanorod magnetophoresis through the extracellular matrix (ECM) via single particle observations in vitro. We show that smaller nanorods experience bimodal stick-slip motion through ECM, as well as larger deviations in their orientation angle with respect to the magnetic field, as compared to larger nanorods

A survey of physical methods for studying nuclear mechanics and mechanobiology

It is increasingly appreciated that the cell nucleus is not only a home for DNA but also a complex material that resists physical deformations and dynamically responds to external mechanical cues. The molecules that confer mechanical properties to nuclei certainly contribute to laminopathies and possibly contribute to cellular mechanotransduction and physical processes in cancer such as metastasis. Studying nuclear mechanics and the downstream biochemical consequences or their modulation requires a suite of complex assays for applying, measuring, and visualizing mechanical forces across diverse length, time, and force scales. Here, we review the current methods in nuclear mechanics and mechanobiology, placing specific emphasis on each of their unique advantages and limitations. Furthermore, we explore important considerations in selecting a new methodology as are demonstrated by recent examples from the literature. We conclude by providing an outlook on the development of new methods and the judicious use of the current techniques for continued exploration into the role of nuclear mechanobiology

Torsional response and stiffening of individual multi-walled carbon nanotubes

We report on the characterization of torsional oscillators which use multi-walled carbon nanotubes as the spring elements. Through atomic-force-microscope force-distance measurements we are able to apply torsional strains to the nanotubes and measure their torsional spring constants and effective shear moduli. We find that the effective shear moduli cover a broad range, with the largest values near the theoretically predicted value. The data also suggest that the nanotubes are stiffened by repeated flexing.Comment: 4 page

Functionalization of carbon nanotubes with proteins and quantum dots in aqueous buffer solutions

We report here on a method of suspending carbon nanotubes (CNTs) in aqueous buffer solutions and functionalizing CNTs with a molecule that is ``sticky'' to proteins. The specific bifunctional molecule used in this study is 1-pyrene butanoic acid succidymidyl ester (1-pbase). We report successful protein and quantum dot functionalization of the CNTs, using 1-pbase as a linking agent

Combined Selective Plane Illumination Microscopy and FRAP Maps Intranuclear Diffusion of NLS-GFP

Since its initial development in 1976, fluorescence recovery after photobleaching (FRAP) has been one of the most popular tools for studying diffusion and protein dynamics in living cells. Its popularity is derived from the widespread availability of confocal microscopes and the relative ease of the experiment and analysis. FRAP, however, is limited in its ability to resolve spatial heterogeneity. Here, we combine selective plane illumination microscopy (SPIM) and FRAP to create SPIM-FRAP, wherein we use a sheet of light to bleach a two-dimensional (2D) plane and subsequently image the recovery of the same image plane. This provides simultaneous quantification of diffusion or protein recovery for every pixel in a given 2D slice, thus moving FRAP measurements beyond these previous limitations. We demonstrate this technique by mapping both intranuclear diffusion of NLS-GFP and recovery of 53BP1-mCherry, a marker for DNA damage, in live MDA-MB-231 cells. SPIM-FRAP proves to be an order of magnitude faster than fluorescence-correlation-spectroscopy-based techniques for such measurements. We observe large length-scale (>∼500 nm) heterogeneity in the recovery times of NLS-GFP, which is validated against simulated data sets. 2D maps of NLS-GFP recovery times showed no pixel-by-pixel correlation with histone density, although slower diffusion was observed in nucleoli. Additionally, recovery of 53BP1-mCherry was observed to be slowed at sites of DNA damage. We finally developed a diffusion simulation for our SPIM-FRAP experiments to compare across techniques. Our measured diffusion coefficients are on the order of previously reported results, thus validating the quantitative accuracy of SPIM-FRAP relative to well-established methods. With the recent rise of accessibility of SPIM systems, SPIM-FRAP is set to provide a straightforward means of quantifying the spatial distribution of protein recovery or diffusion in living cells

In Situ Resistance Measurements of Strained Carbon Nanotubes

We investigate the response of multi-walled carbon nanotubes to mechanical strain applied with an Atomic Force Microscope (AFM) probe. We find that in some samples, changes in the contact resistance dominate the measured resistance change. In others, strain large enough to fracture the tube can be applied without a significant change in the contact resistance. In this case we observe that enough force is applied to break the tube without any change in resistance until the tube fails. We have also manipulated the ends of the broken tube back in contact with each other, re-establishing a finite resistance. We observe that in this broken configuration the resistance of the sample is tunable to values 15-350 kW greater than prior to breaking.Comment: Submitted to Applied Physics Letter

A Model for a Spreading and Melting Droplet on a Heated Substrate

We develop a model to describe the dynamics of a spreading and melting droplet on a heated substrate. The model, developed in the capillary-dominated limit, is geometrical in nature and couples the contact line, trijunction, and phase-change dynamics. The competition between spreading and melting is characterized by a single parameter KT that represents the ratio of the characteristic contact line velocity to the characteristic melting (or phase-change) velocity. A key component of the model is an equation of motion for the solid. This equation of motion, which accounts for global effects through a balance of forces over the entire solid-liquid interface, including capillary effects at the trijunction, acts in a natural way as the trijunction condition. This is in contrast to models of trijunction dynamics during solidification, where it is common to specify a trijunction condition based on local physics alone. The trijunction dynamics, as well as the contact angle, contact line position, and other dynamic quantities for the spreading and melting droplet, are predicted by the model and are compared to an isothermally spreading liquid droplet whose dynamics are controlled exclusively by the contact line. We find that in general the differences between the dynamics of a spreading and melting droplet and that of an isothermally spreading droplet increase as KT increases. We observe that the presence of the solid phase in the spreading and melting configuration tends to inhibit spreading relative to an isothermally spreading droplet of the same initial geometry. Finally, we find that increasing the effect of spreading promotes melting

Lithographically Defined Micropost Arrays for Programmable Actuation and Interfacial Hydrodynamics

Magnetically actuating surface-attached post (ASAP) arrays have great potential in microfluidic flow control, including mixing and pumping. Both passive (nonactuating) and active (actuating) micropillar arrays can also be used to control pressure-driven flow and the motion of microscopic particles carried by the fluid through microfluidic channels. Molding techniques are popular for generating these microstructures. However, fabricating high aspect ratio elastomeric microstructures over large surface areas suffers from practical problems such as damage incurred in the demolding process. Here, we report on a fabrication protocol that generates ASAP with an aspect ratio as high as 23:1 and a cross-sectional area less than 1 μm2 using straightforward photolithography processes. We generated 50 unique ASAP arrays, each occupying an area of 1 mm2 on a silicon mold; these arrays have varied cross-sectional shape and size, aspect ratio, and spacings between neighboring posts. Our protocol also controls the level of magnetic material in the ASAP tips with a centrifugation step. Using a herringbone pattern ASAP array, we have demonstrated control over the relative phase of actuation between neighboring posts. Such ASAP serve as an experimental platform to test current models predicting that reciprocal actuators in close proximity can successfully drive flow in a low Reynolds (Re) number environment

Gearlike rolling motion mediated by commensurate contact: Carbon nanotubes on HOPG

We report on experiments in which multiwall carbon nanotubes (CNT's) are manipulated with atomic force microscopy (AFM) on a graphite highly oriented pyrolytic graphite (HOPG) substrate. We find certain discrete orientations in which the lateral force of manipulation dramatically increases as we rotate the CNT in the plane of the HOPG surface with the AFM tip. The threefold symmetry of these discrete orientations indicates commensurate contact of the hexagonal graphene surfaces of the HOPG and CNT. As the CNT moves into commensurate contact, we observe the motion change from sliding/rotating in-plane to stick-roll motion

Highly responsive core-shell microactuator arrays for use in viscous and viscoelastic fluids

We present a new fabrication method to produce arrays of highly responsive polymer-metal core-shell magnetic microactuators. The core-shell fabrication method decouples the elastic and magnetic structural components such that the actuator response can be optimized by adjusting the core-shell geometry. Our microstructures are 10 μm long, 550 nm in diameter, and electrochemically fabricated in particle track-etched membranes, comprising a poly(dimethylsiloxane) core with a 100 nm Ni shell surrounding the upper 3–8 μm. The structures can achieve deflections of nearly 90° with moderate magnetic fields and are capable of driving fluid flow in a fluid 550 times more viscous than water