Wednesday, April 18, 2018

Electro-optic deflectors deliver advantages over acousto-optical deflectors in a high resolution, ultra-fast force-clamp optical trap

Michael S. Woody, Marco Capitanio, E. Michael Ostap, and Yale E. Goldman

We characterized experimental artifacts arising from the non-linear response of acousto-optical deflectors (AODs) in an ultra-fast force-clamp optical trap and have shown that using electro-optical deflectors (EODs) instead eliminates these artifacts. We give an example of the effects of these artifacts in our ultra-fast force clamp studies of the interaction of myosin with actin filaments. The experimental setup, based on the concept of Capitanio et al. [Nat. Methods 9, 1013–1019 (2012)] utilizes a bead-actin-bead dumbbell held in two force-clamped optical traps which apply a load to the dumbbell to move it at a constant velocity. When myosin binds to actin, the filament motion stops quickly as the total force from the optical traps is transferred to the actomyosin attachment. We found that in our setup, AODs were unsuitable for beam steering due to non-linear variations in beam intensity and deflection angle as a function of driving frequency, likely caused by low-amplitude standing acoustic waves in the deflectors. These aberrations caused instability in the force feedback loops leading to artifactual jumps in the trap position. We demonstrate that beam steering with EODs improves the performance of our instrument. Combining the superior beam-steering capability of the EODs, force acquisition via back-focal-plane interferometry, and dual high-speed FPGA-based feedback loops, we apply precise and constant loads to study the dynamics of interactions between actin and myosin. The same concept applies to studies of other biomolecular interactions.


Optical manipulation of nematic colloids at the interfaces in azo-dye-doped liquid crystals

Andy Ying Guey Fuh, Ming-Hsien Li, Te-Wei Chang, Yu-I Lee, and Shing Trong Wu

This study demonstrates the optical manipulation of colloids dispersed in azo-dye-doped liquid crystals (DDLCs) where the accumulation occurs at the interfaces of the phase domains. We explain the mechanism related to the formation of the domains and the movement of the colloids in DDLCs with respect to the isomerization of azo-dye molecules via the illumination of laser beams. The colloids are dragged to the interfaces of the isotropic/nematic domain and the air bubble/isotropic domain by molecular interaction and Marangoni flows.


Estimation of actomyosin active force maintained by tropomyosin and troponin complex under vertical forces in the in vitro motility assay system

Shuya Ishii, Masataka Kawai, Shin'ichi Ishiwata, Madoka Suzuki

The interaction between actin filaments and myosin molecular motors is a power source of a variety of cellular functions including cell division, cell motility, and muscular contraction. In vitro motility assay examines actin filaments interacting with myosin molecules that are adhered to a substrate (e.g., glass surface). This assay has been the standard method of studying the molecular mechanisms of contraction under an optical microscope. While the force generation has been measured through an optically trapped bead to which an actin filament is attached, a force vector vertical to the glass surface has been largely ignored with the in vitro motility assay. The vertical vector is created by the gap (distance) between the trapped bead and the glass surface. In this report, we propose a method to estimate the angle between the actin filament and the glass surface by optically determining the gap size. This determination requires a motorized stage in a standard epi-fluorescence microscope equipped with optical tweezers. This facile method is applied to force measurements using both pure actin filaments, and thin filaments reconstituted from actin, tropomyosin and troponin. We find that the angle-corrected force per unit filament length in the active condition (pCa = 5.0) decreases as the angle between the filament and the glass surface increases; i.e. as the force in the vertical direction increases. At the same time, we demonstrate that the force on reconstituted thin filaments is approximately 1.5 times larger than that on pure actin filaments. The range of angles we tested was between 11° and 36° with the estimated measurement error less than 6°. These results suggest the ability of cytoplasmic tropomyosin isoforms maintaining actomyosin active force to stabilize cytoskeletal architecture.


Theoretical investigation on asymmetrical spinning and orbiting motions of particles in a tightly focused power-exponent azimuthal-variant vector field

Yingdi Zhang, Yuxiong Xue, Zhuqing Zhu, Guanghao Rui, Yiping Cui, and Bing Gu

We generate a new kind of azimuthal-variant vector field with a distribution of states of polarization (SoPs) described by the square of the azimuthal angle. Owing to asymmetrical SoPs distribution of this localized linearly polarized vector field, the tightly focused field exhibits a double half-moon shaped pattern with the localized elliptical polarization in the cross section of field at the focal plane. Moreover, we study the three-dimensional distributions of spin and orbital linear and angular momenta in the focal region. We numerically investigate the gradient force, radiation force, spin torque, and orbital torque on a dielectric Rayleigh particle produced by the tightly focused vector field. It is found that asymmetrical spinning and orbiting motions of trapped Rayleigh particles can be realized by the use of a tight vector field with power-exponent azimuthal-variant SoPs.


Fundamental Limits of Optical Tweezer Nanoparticle Manipulation Speeds

Jeffrey E. Melzer and Euan McLeod

Optical tweezers are a noncontact method of 3D positioning applicable to the fields of micro- and nanomanipulation and assembly, among others. In these applications, the ability to manipulate particles over relatively long distances at high speed is essential in determining overall process efficiency and throughput. In order to maximize manipulation speeds, it is necessary to increase the trapping laser power, which is often accompanied by undesirable heating effects due to material absorption. As such, the majority of previous studies focus primarily on trapping large dielectric microspheres using slow movement speeds at low laser powers, over relatively short translation distances. In contrast, we push nanoparticle manipulation beyond the region in which maximum lateral movement speed is linearly proportional to laser power, and investigate the fundamental limits imposed by material absorption, thus quantifying maximum possible speeds attainable with optical tweezers. We find that gold and silver nanospheres of diameter 100 nm are limited to manipulation speeds of ∼0.15 mm/s, while polystyrene spheres of diameter 160 nm can reach speeds up to ∼0.17 mm/s, over distances ranging from 0.1 to 1 mm. When the laser power is increased beyond the values used for these maximum manipulation speeds, the nanoparticles are no longer stably trapped in 3D due to weak confinement as a result of material absorption, heating, microbubble formation, and enhanced Brownian motion. We compared this result to our theoretical model, incorporating optical forces in the Rayleigh regime, Stokes’ drag, and absorption effects, and found good agreement. These results show that optical tweezers can be fast enough to compete with other common, serial rapid prototyping and nanofabrication approaches.

Microscopic Engine Powered by Critical Demixing

Falko Schmidt, Alessandro Magazzù, Agnese Callegari, Luca Biancofiore, Frank Cichos, and Giovanni Volpe

We experimentally demonstrate a microscopic engine powered by the local reversible demixing of a critical mixture. We show that, when an absorbing microsphere is optically trapped by a focused laser beam in a subcritical mixture, it is set into rotation around the optical axis of the beam because of the emergence of diffusiophoretic propulsion. This behavior can be controlled by adjusting the optical power, the temperature, and the criticality of the mixture.


Switching of Giant Lateral Force on Sub‐10 nm Particle Using Phase‐Change Nanoantenna

Tun Cao, Jiaxin Bao, Libang Mao

We numerically show that a giant lateral optical force (LOF) acting on sub‐10 nm non‐chiral particles can be obtained using a dipole‐quadrupole (DQ) Fano resonance (FR). This DQ‐FR is excited by an asymmetric plasmonic bowtie nanoantenna array (BNA), which is based on a Au/Ge2Sb2Te5/Au trilayer. The LOF behaves in a direction in which the incident light has neither a spin‐orbit coupling nor any wave propagation. Analytical theory reveals that the LOF originates from the “hotspot” established by the DQ‐FR in the asymmetric BNA. The direction of DQ‐FR induced LOF is reversibly switched with the phase transition of Ge2Sb2Te5, which in turn pushes the achiral nanoparticle sideways in the opposite direction. A photo thermal model is used to study the temporal variation of the temperature of the Ge2Sb2Te5 film to show the potential for transiting the Ge2Sb2Te5 phase. Particularly, the design of hybrid nanostructure integrating a ground metal mirror can excite a strong magnetic resonance, which enhances the DQ‐FR induced LOF and reduces the Brownian motion effect on the nanoparticles. Hence, our scheme leads to a rapid and stable transportation of nanoparticles with radii as small as 10 nm.


Tuesday, April 17, 2018

Sprouting angiogenesis induces significant mechanical heterogeneities and ECM stiffening across length scales in fibrin hydrogels

Benjamin A. Juliar, Mark T. Keating, Yen P. Kong, Elliot L. Botvinick, Andrew J.Putnam

Matrix stiffness is a well-established instructive cue in two-dimensional cell cultures. Its roles in morphogenesis in 3-dimensional (3D) cultures, and the converse effects of cells on the mechanics of their surrounding microenvironment, have been more elusive given the absence of suitable methods to quantify stiffness on a length-scale relevant for individual cell-extracellular matrix (ECM) interactions. In this study, we applied traditional bulk rheology and laser tweezers-based active microrheology to probe mechanics across length scales during the complex multicellular process of capillary morphogenesis in 3D, and further characterized the relative contributions of neovessels and supportive stromal cells to dynamic changes in stiffness over time. Our data show local ECM stiffness was highly heterogeneous around sprouting capillaries, and the variation progressively increased with time. Both endothelial cells and stromal support cells progressively stiffened the ECM, with the changes in bulk properties dominated by the latter. Interestingly, regions with high micro-stiffness did not necessarily correlate with remodeled regions of high ECM density as shown by confocal reflectance microscopy. Collectively, these findings, especially the large spatiotemporal variations in local stiffness around cells during morphogenesis in soft 3D fibrin gels, underscore that characterizing ECM mechanics across length scales. provides an opportunity to attain a deeper mechanobiological understanding of the microenvironment's roles in cell fate and tissue patterning.


Tunable optical forces enhanced by plasmonic modes hybridization in optical trapping of gold nanorods with plasmonic nanocavity

Wen-Hao Huang, Shun-Feng Li, Hai-Tao Xu, Zheng-Xun Xiang, Yong-Bing Long, and Hai-Dong Deng
The optomechanical interaction between a plasmonic nanocavity and a gold nanorod through optical forces is demonstrated. It is revealed that strong localized plasmon resonance mode hybridization induced by a gold nanorod results in the resonance mode of the nanocavity splitting into two different plasmon resonance modes (bonding plasmon resonance mode and antibonding plasmon resonance mode). When the whole system (gold nanorod and gold nanocavity) is excited at the antibonding plasmon mode, the gold nanorod can receive an optical pushing force and be pushed away from the gold nanocavity. On the other hand, an optical pulling force acts on the gold nanorod and the gold nanorod can be trapped by the gold nanocavity when the plasmonic tweezers work at the bonding mode. The optical pulling force acting on the gold nanorod can be enhanced by two orders of magnitude larger than that of the same sized dielectric nanorod, which benefits from the strong resonant nearfield interaction between the gold nanorod and the gold nanocavity. More importantly, the shape and the position of the optical potential can be tuned by tailoring the wavelength of the laser used in the optical trapping, which can be used to manipulate the gold nanorod within a nanoscale region. Our findings have important implications for optical trapping, manipulation, sorting, and sieving of plasmonic nanoparticles with plasmonic tweezers.


A cooperative transition from the semi-flexible to the flexible regime of polymer elasticity: Mitoxantrone-induced DNA condensation

C.H.M.Lima, G.O.Almeida, M.S.Rocha

We report a high cooperative transition from the semi-flexible to the flexible regime of polymer elasticity during the interaction of the DNA molecule with the chemotherapeutic drug Mitoxantrone (MTX). By using single molecule force spectroscopy, we show that the force-extension curves of the DNA-MTX complexes deviate from the typical worm-like chain behavior as the MTX concentration in the sample increases, becoming straight lines for sufficiently high drug concentrations. The behavior of the radius of gyration of the complexes as a function of the bound MTX concentration was used to quantitatively investigate the cooperativity of the condensation process. The present methodology can be promptly applied to other ligands that condense the DNA molecule upon binding, opening new possibilities in the investigation of this type of process and, more generally, in the investigation of phase transitions in polymer physics.