Jennifer E. Curtis

Dynamic holographic optical tweezers

Optical trapping is an increasingly important technique for controlling and probing matter at length scales ranging from nanometers to millimeters. This paper describes methods for creating large numbers of high-quality optical traps in arbitrary three-dimensional configurations and for dynamically reconfiguring them under computer control. In addition to forming conventional optical tweezers, these methods also can sculpt the wavefront of each trap individually, allowing for mixed arrays of traps based on different modes of light, including optical vortices, axial line traps, optical bottles and optical rotators. The ability to establish large numbers of individually structured optical traps and to move them independently in three dimensions promises exciting new opportunities for research, engineering, diagnostics, and manufacturing at mesoscopic lengthscales.

How vinculin regulates force transmission

Focal adhesions mediate force transfer between ECM-integrin complexes and the cytoskeleton. Although vinculin has been implicated in force transmission, few direct measurements have been made, and there is little mechanistic insight. Using vinculin-null cells expressing vinculin mutants, we demonstrate that vinculin is not required for transmission of adhesive and traction forces but is necessary for myosin contractility-dependent adhesion strength and traction force and for the coupling of cell area and traction force. Adhesion strength and traction forces depend differentially on vinculin head (VH) and tail domains. VH enhances adhesion strength by increasing ECM-bound integrin–talin complexes, independently from interactions with vinculin tail ligands and contractility. A full-length, autoinhibition-deficient mutant (T12) increases adhesion strength compared with VH, implying roles for both vinculin activation and the actin-binding tail. In contrast to adhesion strength, vinculin-dependent traction forces absolutely require a full-length and activated molecule; VH has no effect. Physical linkage of the head and tail domains is required for maximal force responses. Residence times of vinculin in focal adhesions, but not T12 or VH, correlate with applied force, supporting a mechanosensitive model for vinculin activation in which forces stabilize vinculin’s active conformation to promote force transfer.

Modulated optical vortices.

Single-beam optical gradient force traps created by focusing helical modes of light are known as optical vortices. Modulating the helical pitch of such a modes wave front yields a new class of optical traps whose dynamically reconfigurable intensity distributions provide new opportunities for controlling motion in mesoscopic systems. An implementation of modulated optical vortices based on the dynamic holographic optical tweezer technique is described.

Symmetry dependence of holograms for optical trapping.

No iterative algorithm is necessary to calculate holograms for most holographic optical trapping patterns. Instead, holograms may be produced by a simple extension of the prisms-and-lenses method. This formulaic approach yields the same diffraction efficiency as iterative algorithms for any asymmetric or symmetric but nonperiodic pattern of points while requiring less calculation time. A slight spatial disordering of periodic patterns significantly reduces intensity variations between the different traps without extra calculation costs. Eliminating laborious hologram calculations should greatly facilitate interactive holographic trapping.

Nonperturbative Chemical Modification of Graphene for Protein Micropatterning

Graphenes extraordinary physical properties and its planar geometry make it an ideal candidate for a wide array of applications, many of which require controlled chemical modification and the spatial organization of molecules on its surface. In particular, the ability to functionalize and micropattern graphene with proteins is relevant to bioscience applications such as biomolecular sensors, single-cell sensors, and tissue engineering. We report a general strategy for the noncovalent chemical modification of epitaxial graphene for protein immobilization and micropatterning. We show that bifunctional molecule pyrenebutanoic acid-succinimidyl ester (PYR-NHS), composed of the hydrophobic pyrene and the reactive succinimide ester group, binds to graphene noncovalently but irreversibly. We investigate whether the chemical treatment perturbs the electronic band structure of graphene using X-ray photoemission (XPS) and Raman spectroscopy. Our results show that the sp(2) hybridization remains intact and that the π band maintains its characteristic Lorentzian shape in the Raman spectra. The modified graphene surfaces, which bind specifically to amines in proteins, are micropatterned with arrays of fluorescently labeled proteins that are relevant to glucose sensors (glucose oxidase) and cell sensor and tissue engineering applications (laminin).

Tuning the orbital angular momentum in optical vortex beams

We introduce a method to tune the local orbital angular momentum density in an optical vortex beam without changing its topological charge or geometric intensity distribution. We show that adjusting the relative amplitudes a and b of two interfering collinear vortex beams of equal but opposite helicity provides the smooth variation of the orbital angular momentum density in the resultant vortex beam. Despite the azimuthal intensity modulations that arise from the interference, the local orbital angular momentum remains constant on the vortex annulus and scales with the modulation parameter, c = (a-b)/(a+b).

An in-tumor genetic screen reveals that the BET bromodomain protein, BRD4, is a potential therapeutic target in ovarian carcinoma.

Significance The observations presented here demonstrate that inhibition of the BET bromodomain protein, BRD4, is a potential therapeutic approach to high-grade epithelial ovarian cancers that exhibit elevated MYCN expression. As BRD4 inhibitors enter clinical studies, these findings provide a rationale for stratification of patients in whom to test the effects of BRD4 inhibition. High-grade serous ovarian carcinoma (HGSOC) is the most common and aggressive form of epithelial ovarian cancer, for which few targeted therapies exist. To search for new therapeutic target proteins, we performed an in vivo shRNA screen using an established human HGSOC cell line growing either subcutaneously or intraperitoneally in immunocompromised mice. We identified genes previously implicated in ovarian cancer such as AURKA1, ERBB3, CDK2, and mTOR, as well as several novel candidates including BRD4, VRK1, and GALK2. We confirmed, using both genetic and pharmacologic approaches, that the activity of BRD4, an epigenetic transcription modulator, is necessary for proliferation/survival of both an established human ovarian cancer cell line (OVCAR8) and a subset of primary serous ovarian cancer cell strains (DFs). Among the DFs tested, the strains sensitive to BRD4 inhibition revealed elevated expression of either MYCN or c-MYC, with MYCN expression correlating closely with JQ1 sensitivity. Accordingly, primary human xenografts derived from high-MYCN or c-MYC strains exhibited sensitivity to BRD4 inhibition. These data suggest that BRD4 inhibition represents a new therapeutic approach for MYC-overexpressing HGSOCs.

High-precision steering of multiple holographic optical traps

Locating and steering entire ensembles of microscopic objects has become extremely practical with the emergence of holographic optical tweezers. Application of this technology to single molecule experiments requires great accuracy in the spatial positioning of optical traps. This paper calculates the theoretical position resolution of a single holographic beam, predicting that sub-nanometer resolution is easily achieved. Experimental corroboration of the spatial resolutions inverse dependence on the holograms number of pixels and phase levels is presented. To at least a nanometer range position resolution, multiple optical tweezers created by complex superposition holograms also follow the theoretical predictions for a single beam.

Nanopatterning reconfigurable magnetic landscapes via thermally assisted scanning probe lithography

The search for novel tools to control magnetism at the nanoscale is crucial for the development of new paradigms in optics, electronics and spintronics. So far, the fabrication of magnetic nanostructures has been achieved mainly through irreversible structural or chemical modifications. Here, we propose a new concept for creating reconfigurable magnetic nanopatterns by crafting, at the nanoscale, the magnetic anisotropy landscape of a ferromagnetic layer exchange-coupled to an antiferromagnetic layer. By performing localized field cooling with the hot tip of a scanning probe microscope, magnetic structures, with arbitrarily oriented magnetization and tunable unidirectional anisotropy, are reversibly patterned without modifying the film chemistry and topography. This opens unforeseen possibilities for the development of novel metamaterials with finely tuned magnetic properties, such as reconfigurable magneto-plasmonic and magnonic crystals. In this context, we experimentally demonstrate spatially controlled spin wave excitation and propagation in magnetic structures patterned with the proposed method.

Optical force sensor array in a microfluidic device based on holographic optical tweezers

Holographic optical tweezers (HOT) are a versatile technology, with which complex arrays and movements of optical traps can be realized to manipulate multiple microparticles in parallel and to measure the forces affecting them in the piconewton range. We report on the combination of HOT with a fluorescence microscope and a stop-flow, multi-channel microfluidic device. The integration of a high-speed camera into the setup allows for the calibration of all the traps simultaneously both using Boltzmann statistics or the power spectrum density of the particle diffusion within the optical traps. This setup permits complete spatial, chemical and visual control of the microenvironment applicable to probing chemo-mechanical properties of cellular or subcellular structures. As an example we constructed a biomimetic, quasi-two-dimensional actin network on an array of trapped polystyrene microspheres inside the microfluidic chamber. During crosslinking of the actin filaments by Mg(2+) ions, we observe the build up of mechanical tension throughout the actin network. Thus, we demonstrate how our integrated HOT-microfluidics platform can be used as a reconfigurable force sensor array with piconewton resolution to investigate chemo-mechanical processes.