Friday, January 27, 2017

α-Actinin/titin interaction: A dynamic and mechanically stable cluster of bonds in the muscle Z-disk

Marco Grison, Ulrich Merkel, Julius Kostan, Kristina Djinović-Carugo, and Matthias Rief

Stable anchoring of titin within the muscle Z-disk is essential for preserving muscle integrity during passive stretching. One of the main candidates for anchoring titin in the Z-disk is the actin cross-linker α-actinin. The calmodulin-like domain of α-actinin binds to the Z-repeats of titin. However, the mechanical and kinetic properties of this important interaction are still unknown. Here, we use a dual-beam optical tweezers assay to study the mechanics of this interaction at the single-molecule level. A single interaction of α-actinin and titin turns out to be surprisingly weak if force is applied. Depending on the direction of force application, the unbinding forces can more than triple. Our results suggest a model where multiple α-actinin/Z-repeat interactions cooperate to ensure long-term stable titin anchoring while allowing the individual components to exchange dynamically.


Optical sorting and cultivation of denitrifying anaerobic methane oxidation archaea

Xiaoqiong Qi, David M. Carberry, Chen Cai, Shihu Hu, Zhiguo Yuan, Halin Rubinsztein Dunlop, and Jianhua Guo

Denitrifying anaerobic methane oxidizing (DAMO) microorganisms play an important role in the global carbon and nitrogen cycles as they are able to mediate methane oxidation using nitrite/nitrate under anoxic conditions. However, the physiological properties of DAMO microorganisms remain poorly understood, partially since the organisms are difficult to isolate or cultivate in pure culture and partially because of their long cultivation time. In this study, DAMO cell sorting has been conducted by integrating optical tweezers within enclosed microfluidic chips. This integrated cell sorting method has high purity, low infection rates, and causes no discernable harm to cell viability. The purity of the sorted cells was controlled by the microfluidic chip structure design and operation, while the cell viability was verified by imaging the cultured DAMO archaea after 420 days.


Optical manipulation for studies of collisional dynamics of micron-sized droplets under gravity

Maksym Ivanov, Kelken Chang, Ivan Galinskiy, Bernhard Mehlig, and Dag Hanstorp

A new experimental technique for creating and imaging collisions of micron-sized droplets settling under gravity is presented. A pair of glycerol droplets is suspended in air by means of two optical traps. The droplet relative velocities are determined by the droplet sizes. The impact parameter is precisely controlled by positioning the droplets using the two optical traps. The droplets are released by turning off the trapping light using electro-optical modulators. The motion of the sedimenting droplets is then captured by two synchronized high-speed cameras, at a frame rate of up to 63 kHz. The method allows the direct imaging of the collision of droplets without the influence of the optical confinement imposed by the trapping force. The method will facilitate efficient studies of the microphysics of neutral, as well as charged, liquid droplets and their interactions with light, electric field and thermodynamic environment, such as temperature or vapor concentration.


A Clostridium difficile-Specific, Gel-Forming Protein Required for Optimal Spore Germination

M. Lauren Donnelly, William Li, Yong-qing Li, Lauren Hinkel, Peter Setlow, Aimee Shen

Clostridium difficile is a Gram-positive spore-forming obligate anaerobe that is a leading cause of antibiotic-associated diarrhea worldwide. In order for C. difficile to initiate infection, its aerotolerant spore form must germinate in the gut of mammalian hosts. While almost all spore-forming organisms use transmembrane germinant receptors to trigger germination, C. difficile uses the pseudoprotease CspC to sense bile salt germinants. CspC activates the related subtilisin-like protease CspB, which then proteolytically activates the cortex hydrolase SleC. Activated SleC degrades the protective spore cortex layer, a step that is essential for germination to proceed. Since CspC incorporation into spores also depends on CspA, a related pseudoprotease domain, Csp family proteins play a critical role in germination. However, how Csps are incorporated into spores remains unknown. In this study, we demonstrate that incorporation of the CspC, CspB, and CspA germination regulators into spores depends on CD0311 (renamed GerG), a previously uncharacterized hypothetical protein. The reduced levels of Csps in gerG spores correlate with reduced responsiveness to bile salt germinants and increased germination heterogeneity in single-spore germination assays. Interestingly, asparagine-rich repeat sequences in GerG’s central region facilitate spontaneous gel formation in vitro even though they are dispensable for GerG-mediated control of germination. Since GerG is found exclusively in C. difficile, our results suggest that exploiting GerG function could represent a promising avenue for developing C. difficile-specific anti-infective therapies.


Force generation by titin folding

Zsolt Mártonfalvi, Pasquale Bianco, Katalin Naftz, György G. Ferenczy, Miklós Kellermayer

Titin is a giant protein that provides elasticity to muscle. As the sarcomere is stretched, titin extends hierarchically according to the mechanics of its segments. Whether titin's globular domains unfold during this process and how such unfolded domains might contribute to muscle contractility are strongly debated. To explore the force-dependent folding mechanisms, here we manipulated skeletal-muscle titin molecules with high-resolution optical tweezers. In force-clamp mode, after quenching the force (<10 pN), extension fluctuated without resolvable discrete events. In position-clamp experiments the time-dependent force trace contained rapid fluctuations and a gradual increase of average force, indicating that titin can develop force via dynamic transitions between its structural states en route to the native conformation. In 4 M urea, which destabilizes H-bonds hence the consolidated native domain structure, the net force increase disappeared but the fluctuations persisted. Thus, whereas net force generation is caused by the ensemble folding of the elastically-coupled domains, force fluctuations arise due to a dynamic equilibrium between unfolded and molten-globule states. Monte-Carlo simulations incorporating a compact molten-globule intermediate in the folding landscape recovered all features of our nanomechanics results. The ensemble molten-globule dynamics delivers significant added contractility that may assist sarcomere mechanics, and it may reduce the dissipative energy loss associated with titin unfolding/refolding during muscle contraction/relaxation cycles.


Thursday, January 26, 2017

Single Upconversion Nanoparticle–Bacterium Cotrapping for Single-Bacterium Labeling and Analysis

Hongbao Xin, Yuchao Li, Dekang Xu, Yueli Zhang, Chia-Hung Chen, Baojun Li

Detecting and analyzing pathogenic bacteria in an effective and reliable manner is crucial for the diagnosis of acute bacterial infection and initial antibiotic therapy. However, the precise labeling and analysis of bacteria at the single-bacterium level are a technical challenge but very important to reveal important details about the heterogeneity of cells and responds to environment. This study demonstrates an optical strategy for single-bacterium labeling and analysis by the cotrapping of single upconversion nanoparticles (UCNPs) and bacteria together. A single UCNP with an average size of ≈120 nm is first optically trapped. Both ends of a single bacterium are then trapped and labeled with single UCNPs emitting green light. The labeled bacterium can be flexibly moved to designated locations for further analysis. Signals from bacteria of different sizes are detected in real time for single-bacterium analysis. This cotrapping method provides a new approach for single-pathogenic-bacterium labeling, detection, and real-time analysis at the single-particle and single-bacterium level.

Energetics, kinetics, and pathway of SNARE folding and assembly revealed by optical tweezers

Yongli Zhang

Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) are universal molecular engines that drive membrane fusion. Particularly, synaptic SNAREs mediate fast calcium-triggered fusion of neurotransmitter-containing vesicles with plasma membranes for synaptic transmission, the basis of all thought and action. During membrane fusion, complementary SNAREs located on two apposed membranes (often called t- and v-SNAREs) join together to assemble into a parallel four-helix bundle, releasing the energy to overcome the energy barrier for fusion. A long-standing hypothesis suggests that SNAREs act like a zipper to draw the two membranes into proximity and thereby force them to fuse. However, a quantitative test of this SNARE zippering hypothesis was hindered by difficulties to determine the energetics and kinetics of SNARE assembly and to identify the relevant folding intermediates. Here, we first review different approaches that have been applied to study SNARE assembly and then focus on high-resolution optical tweezers (OTs). We summarize the folding energies, kinetics, and pathways of both wild-type and mutant SNARE complexes derived from this new approach. These results show that synaptic SNAREs assemble in four distinct stages with different functions: slow N-terminal domain (NTD) association initiates SNARE assembly; a middle domain (MD) suspends and controls SNARE assembly; and rapid sequential zippering of the C-terminal domain (CTD) and the linker domain (LD) directly drive membrane fusion. In addition, the kinetics and pathway of the stage-wise assembly are shared by other SNARE complexes. These measurements prove the SNARE zippering hypothesis and suggest new mechanisms for SNARE assembly regulated by other proteins.

The frequency-dependent response of single aerosol particles to vapour phase oscillations and its application in measuring diffusion coefficients

Thomas C. Preston, James F. Davies and Kevin R. Wilson

A new method for measuring diffusion in the condensed phase of single aerosol particles is proposed and demonstrated. The technique is based on the frequency-dependent response of a binary particle to oscillations in the vapour phase of one of its chemical components. We discuss how this physical situation allows for what would typically be a non-linear boundary value problem to be approximately reduced to a linear boundary value problem. For the case of aqueous aerosol particles, we investigate the accuracy of the closed-form analytical solution to this linear problem through a comparison with the numerical solution of the full problem. Then, using experimentally measured whispering gallery modes to track the frequency-dependent response of aqueous particles to relative humidity oscillations, we determine diffusion coefficients as a function of water activity. The measured diffusion coefficients are compared to previously reported values found using the two common experiments: (i) the analysis of the sorption/desorption of water from a particle after a step-wise change to the surrounding relative humidity and (ii) the isotopic exchange of water between a particle and the vapour phase. The technique presented here has two main strengths: first, when compared to the sorption/desorption experiment, it does not require the numerical evaluation of a boundary value problem during the fitting process as a closed-form expression is available. Second, when compared to the isotope exchange experiment, it does not require the use of labeled molecules. Therefore, the frequency-dependent experiment retains the advantages of these two commonly used methods but does not suffer from their drawbacks.


Rigorous analysis of optical forces between two dielectric planar waveguides immersed in dielectric fluid media

Janderson R. Rodrigues, Vilson R. Almeida

This work presents a rigorous analysis of optical forces between planar waveguides immersed in an arbitrary background medium. This approach exploits the Minkowski stress tensor formulation, which is compared with a normalized version of the dispersion relation method, showing excellent results agreement for different dielectric fluid media. Due to slot-waveguide effect, optical forces from TM modes are more sensitive to changes in the fluid refractive index than the TE counterparts. Furthermore, the repulsive optical force from the antisymmetric TM1 mode becomes stronger for higher refractive indexes, whereas the attractive force of the symmetric TM0 mode becomes weaker. The methodology and results presented in this work provide a rigorous analysis of nano-optomechanical devices actuated by optical forces in a broad range of materials and applications. Therefore, this study may impact areas of light-induced interactions presenting novel optofluidic and optomechanical functionalities, thus finding applications in nanoscale transport,


Rotation and deformation of human red blood cells with light from tapered fiber probes

Xiaoshuai Liu, Jianbin Huang, Yuchao Li, Yao Zhang, Baojun Li

Dynamic rotation and deformation of human red blood cells (RBCs) are extremely important to investigate the survival and mechanical features of cells, which will be of great physiological and pathological significance. Here, we report an optical approach that is capable of both rotating and deforming RBCs with light from two tapered fiber probes (TFPs). With laser beams at the wavelength of 980 nm injected into the TFPs, a single RBC was rotated around different axes while single or multiple RBCs were stretched by adjusting the points of action and magnitude of the optical forces from the TFPs. The biological safety of the approach was also discussed by taking the laser power required into account.


Wednesday, January 25, 2017

Optical Trapping Nanometry of Hypermethylated CPG-Island DNA

Csaba I. Pongor, Pasquale Bianco, György Ferenczy, Richárd Kellermayer, Miklós Kellermayer

Cytosine methylation is a key mechanism of epigenetic regulation. CpG-dense loci, called “CpG islands”, play a particularly important role in modulating gene expression. Methylation has long been suspected to alter the physical properties of DNA, but the full spectrum of the evoked changes is unknown. Here we measured the methylation-induced nanomechanical changes in a DNA molecule with the sequence of a CpG island. For the molecule under tension, contour length, bending rigidity and intrinsic stiffness decreased in hypermethylated dsDNA, pointing at structural compaction which may facilitate DNA packaging in vivo. Intriguingly, increased forces were required to convert hypermethylated dsDNA into an extended S-form configuration. The reduction of force hysteresis during mechanical relaxation indicated that methylation generates a barrier against strand unpeeling and melting-bubble formation. The high structural stability is likely to have significant consequences on the recognition, replication, transcription, and reparation of hypermethylated genetic regions.


In situ calibration of position detection in an optical trap for active microrheology in viscous materials

Jack R. Staunton, Ben Blehm, Alexus Devine, and Kandice Tanner

In optical trapping, accurate determination of forces requires calibration of the position sensitivity relating displacements to the detector readout via the V-nm conversion factor (β). Inaccuracies in measured trap stiffness (k) and dependent calculations of forces and material properties occur if β is assumed to be constant in optically heterogeneous materials such as tissue, necessitating calibration at each probe. For solid-like samples in which probes are securely positioned, calibration can be achieved by moving the sample with a nanopositioning stage and stepping the probe through the detection beam. However, this method may be applied to samples only under select circumstances. Here, we introduce a simple method to find β in any material by steering the detection laser beam while the probe is trapped. We demonstrate the approach in the yolk of living Danio rerio (zebrafish) embryos and measure the viscoelastic properties over an order of magnitude of stress-strain amplitude.


Acoustical and optical radiation pressures and the development of single beam acoustical tweezers

Jean-Louis Thomas, Régis Marchiano, Diego Baresch

Studies on radiation pressure in acoustics and optics have enriched one another and have a long common history. Acoustic radiation pressure is used for metrology, levitation, particle trapping and actuation. However, the dexterity and selectivity of single-beam optical tweezers are still to be matched with acoustical devices. Optical tweezers can trap, move and position micron size particles, biological samples or even atoms with subnanometer accuracy in three dimensions. One limitation of optical tweezers is the weak force that can be applied without thermal damage due to optical absorption. Acoustical tweezers overcome this limitation since the radiation pressure scales as the field intensity divided by the speed of propagation of the wave. However, the feasibility of single beam acoustical tweezers was demonstrated only recently. In this paper, we propose a historical review of the strong similarities but also the specificities of acoustical and optical radiation pressures, from the expression of the force to the development of single-beam acoustical tweezers.


Plasmonic non-concentric nanorings array as an unidirectional nano-optical conveyor belt actuated by polarization rotation

Min Jiang, Guanghui Wang, Wenxiang Jiao, Zhoufeng Ying, Ningmu Zou, Ho-pui Ho, Tianyu Sun, and Xuping Zhang

We report a nano-optical conveyor belt containing an array of gold plasmonic non-concentric nanorings (PNNRs) for the realization of trapping and unidirectional transportation of nanoparticles through rotating the polarization of an excitation beam. The location of hot spots within an asymmetric plasmonic nanostructure is polarization dependent, thus making it possible to manipulate a trapped target by rotating the incident polarization state. In the case of PNNR, the two poles have highly unbalanced trap potential. This greatly enhances the chance of transferring trapped particles between adjacent PNNRs in a given direction through rotating the polarization. As confirmed by three-dimensional finite-difference time-domain analysis, an array of PNNRs forms an unidirectional nano-optical conveyor belt, which delivers target nanoparticles or biomolecules over a long distance with nanometer accuracy. With the capacity to trap and to transfer, our design offers a versatile scheme for conducting mechanical sample manipulation in many on-chip optofluidic applications.


Photoinduced Force Mapping of Plasmonic Nanostructures

Thejaswi U. Tumkur, Xiao Yang, Benjamin Cerjan, Naomi J. Halas, Peter Nordlander, and Isabell Thomann

The ability to image the optical near-fields of nanoscale structures, map their morphology, and concurrently obtain spectroscopic information, all with high spatiotemporal resolution, is a highly sought-after technique in nanophotonics. As a step toward this goal, we demonstrate the mapping of electromagnetic forces between a nanoscale tip and an optically excited sample consisting of plasmonic nanostructures with an imaging platform based on atomic force microscopy. We present the first detailed joint experimental–theoretical study of this type of photoinduced force microscopy. We show that the enhancement of near-field optical forces in gold disk dimers and nanorods follows the expected plasmonic field enhancements with strong polarization sensitivity. We then introduce a new way to evaluate optically induced tip–sample forces by simulating realistic geometries of the tip and sample. We decompose the calculated forces into in-plane and out-of-plane components and compare the calculated and measured force enhancements in the fabricated plasmonic structures. Finally, we show the usefulness of photoinduced force mapping for characterizing the heterogeneity of near-field enhancements in precisely e-beam fabricated nominally alike nanostructures - a capability of widespread interest for precise nanomanufacturing, SERS, and photocatalysis applications.

Tuesday, January 24, 2017

Pre-T Cell Receptors (Pre-TCRs) Leverage Vβ Complementarity Determining Regions (CDRs) and Hydrophobic Patch in Mechanosensing Thymic Self-ligands

Dibyendu Kumar Das, Robert J. Mallis, Jonathan S. Duke-Cohan, Rebecca E. Hussey, Paul W. Tetteh, Mark Hilton, Gerhard Wagner, Matthew J. Lang and Ellis L. Reinherz

The pre-T cell receptor (pre-TCR) is a pTα-β heterodimer functioning in early αβ T cell development. Although once thought to be ligand-autonomous, recent studies show that pre-TCRs participate in thymic repertoire formation through recognition of peptides bound to major histocompatibility molecules (pMHC). Using optical tweezers, we probe pre-TCR bonding with pMHC at the single molecule level. Like the αβTCR, the pre-TCR is a mechanosensor undergoing force-based structural transitions that dynamically enhance bond lifetimes and exploiting allosteric control regulated via the Cβ FG loop region. The pre-TCR structural transitions exhibit greater reversibility than TCRαβ and ordered force-bond lifetime curves. Higher piconewton force requires binding through both complementarity determining region loops and hydrophobic Vβ patch apposition. This patch functions in the pre-TCR as a surrogate Vα domain, fostering ligand promiscuity to favor development of β chains with self-reactivity but is occluded by α subunit replacement of pTα upon αβTCR formation. At the double negative 3 thymocyte stage where the pre-TCR is first expressed, pre-TCR interaction with self-pMHC ligands imparts growth and survival advantages as revealed in thymic stromal cultures, imprinting fundamental self-reactivity in the T cell repertoire. Collectively, our data imply the existence of sequential mechanosensor αβTCR repertoire tuning via the pre-TCR.


Vertical oscillations of dust particles in a strongly magnetized plasma sheath induced by horizontal laser manipulation

M. Puttscher, A. Melzer, U. Konopka, S. LeBlanc, B. Lynch, and E. Thomas Jr.

Experimental studies are presented where dust particles are suspended in the lower sheath region of an argon rf discharge at a strong vertical magnetic field from B=1.5B=1.5 T up to 2.272.27 T. There the particles arranged in an ordered pattern imposed by the upper mesh electrode. It is observed that the particles jump to a new equilibrium position, where they exhibit self-excited vertical oscillations when illuminated by a horizontal laser beam. The dust motion is weakly damped during an upward jump and strongly damped during the return to the equilibrium after the laser is switched off. A model based on delayed charging is presented that can describe the observed behavior.

Optical Trap-Mediated High-Sensitivity Nanohole Array Biosensors with Random Nanospikes

Takayasu Yoshikawa, Mamoru Tamura, Shiho Tokonami, and Takuya Iida

We clarify an unconventional principle of the light-driven operation of a biosensor for enhanced sensitivity with the help of random nanospikes added to the surface of a nanohole array. Such a system is capable of optically guiding viruses and trapping them in the vicinity of a highly sensitive site by an anomalous light-induced force arising from random-nanospike-modulated extraordinary optical transmission and the plasmonic mirror image in a virus as a dielectric submicron object. In particular, after guiding the viruses near the apex of nanospikes, there are conditions where the spectral peak shift of extraordinary optical transmission can be greatly increased and reach several hundred nanometers in comparison with that of a conventional nanohole array without random nanospikes. These results will allow for the development of a simple, rapid, and highly sensitive virus detection method based on optical trapping with the help of random-nanospike-modulated extraordinary optical transmission, facilitating convenient medical diagnosis and food inspection.


Enhanced optical magnetism for reversed optical binding forces between silicon nanoparticles in the visible region

Taka-aki Yano, Yuta Tsuchimoto, Remo Proietti Zaccaria, Andrea Toma, Alejandro Portela, and Masahiko Hara

We perform a comprehensive numerical analysis on the optical binding forces of a multiple-resonant silicon nanodimer induced by the normal illumination of a plane wave in the visible region. The silicon nanodimer provides either repulsive or attractive forces in water while providing only attractive forces in air. The enhancement of the magnetic dipole mode is attributed to the generation of repulsive forces. The sign (attractive/repulsive) and the amplitude of the optical forces are controlled by incident polarization and separation distance between the silicon nanoparticles. These optomechanical effects demonstrate a key step toward the optical sorting and assembly of silicon nanoparticles.


Ponderomotive convection in water induced by a CW laser

M. N. Shneider and V. V. Semak

An optically induced convection during IR laser interaction with water or any absorbing liquid is described theoretically. The numerical simulations performed using the developed model show that the optical pressure and ponderomotive forces produce water flow in the direction of the laser beam propagation. In the later stage of interaction, when the water temperature rises, the Archimedes force becomes comparable and, ultimately, dominant, producing convection directed against the vector of gravitational acceleration (upward). The theoretical estimates and numerical simulations predict fluid dynamics similar to what is observed in previous experiments.


Monday, January 23, 2017

A Cellular Model of Shear-Induced Hemolysis

Salman Sohrabi, Yaling Liu

A novel model is presented to study red blood cell (RBC) hemolysis at cellular level. Under high shear rates, pores form on RBC membranes through which hemoglobin (Hb) leaks out and increases free Hb content of plasma leading to hemolysis. By coupling lattice Boltzmann and spring connected network models through immersed boundary method, we estimate hemolysis of a single RBC under various shear rates. First, we use adaptive meshing to find local strain distribution and critical sites on RBC membranes, and then we apply underlying molecular dynamics simulations to evaluate damage. Our approach comprises three sub-models: defining criteria of pore formation, calculating pore size, and measuring Hb diffusive flux out of pores. Our damage model uses information of different scales to predict cellular level hemolysis. Results are compared with experimental studies and other models in literature. The developed cellular damage model can be used as a predictive tool for hydrodynamic and hematologic design optimization of blood-wetting medical devices.


Photothermal heating of nanoribbons

Bennett E. Smith ; Xuezhe Zhou ; E. James Davis ; Peter J. Pauzauskie

Nanoscale optical materials are of great interest for building future optoelectronic devices for information processing and sensing applications. Although heat transfer ultimately limits the maximum power at which nanoscale devices may operate, gaining a quantitative experimental measurement of photothermal heating within single nanostructures remains a challenge. Here, we measure the nonlinear optical absorption coefficient of optically trapped cadmium-sulfide nanoribbons at the level of single nanostructures through observations of their Brownian dynamics during single-beam laser trapping experiments. A general solution to the heat transfer partial differential equation is derived for nanostructures having rectilinear morphology including nanocubes and nanoribbons. Numerical electromagnetic calculations using the discrete-dipole approximation enable the simulation of the photothermal heating source function and the extraction of nonlinear optical absorption coefficients from experimental observations of single nanoribbon dynamics.


Influencing colloidal formation with optical traps

Ifat Jacob, Eitan Edri, Erel Lasnoy, Silvia Piperno and Hagay Shpaisman

We present a novel concept where optical traps are used to influence an ongoing polymerization process of emulsion droplets. By directed coalescence and partial fusion of intermediate nucleation sites, spherical and elongated colloids with specific dimensions are formed. The strength of this approach lies in its versatility and ease of making various changes to the end product without the need for chemical modifications.


Fractal Conical Lens Optical Tweezers

Zhirong Liu; Philip H. Jones

We propose a novel optical tweezers composed of an annular beam with alternate radially and azimuthally polarized rings modulated by a fractal conical lens (FCL) and demonstrate its optical forces on Rayleigh dielectric particles both analytically and numerically. Owing to the optical system's particular focusing properties, which could generate a dark-centered or peak-centered intensity distribution in the focal region when selecting an appropriate truncation parameter in front of the focusing lens, the proposed FCL optical tweezers could selectively trap and manipulate dielectric mesoscopic particles with low- or high-refractive indices by appropriately adjusting the radius of the pupil or the beam. Finally, the stability conditions for effective trapping and manipulation Rayleigh particles are analyzed.


Thursday, January 19, 2017

Rotation of single live mammalian cells using dynamic holographic optical tweezers

Bin Cao, Laimonas Kelbauska, Samantha Chan, Rishabh M. Shetty, Dean Smith, Deirdre R. Meldrum

We report on a method for rotating single mammalian cells about an axis perpendicular to the optical system axis through the imaging plane using dynamic holographic optical tweezers (HOTs). Two optical traps are created on the opposite edges of a mammalian cell and are continuously transitioned through the imaging plane along the circumference of the cell in opposite directions, thus providing the torque to rotate the cell in a controlled fashion. The method enables a complete 360° rotation of live single mammalian cells with spherical or near-to spherical shape in 3D space, and represents a useful tool suitable for the single cell analysis field, including tomographic imaging.


Mapping intracellular mechanics on micropatterned substrates

Kalpana Mandal, Atef Asnacios, Bruno Goud, and Jean-Baptiste Manneville

The mechanical properties of cells impact on their architecture, their migration, intracellular trafficking, and many other cellular functions and have been shown to be modified during cancer progression. We have developed an approach to map the intracellular mechanical properties of living cells by combining micropatterning and optical tweezers-based active microrheology. We optically trap micrometer-sized beads internalized in cells plated on crossbow-shaped adhesive micropatterns and track their displacement following a step displacement of the cell. The local intracellular complex shear modulus is measured from the relaxation of the bead position assuming that the intracellular microenvironment of the bead obeys power-law rheology. We also analyze the data with a standard viscoelastic model and compare with the power-law approach. We show that the shear modulus decreases from the cell center to the periphery and from the cell rear to the front along the polarity axis of the micropattern. We use a variety of inhibitors to quantify the spatial contribution of the cytoskeleton, intracellular membranes, and ATP-dependent active forces to intracellular mechanics and apply our technique to differentiate normal and cancer cells.


Extraordinary Optical Transmission: Fundamentals and Applications

Sergio G. Rodrigo; Fernando de León-Pérez; Luis Martín-Moreno

Extraordinary optical transmission (EOT) is a term that refers to electromagnetic resonances through sets of subwavelength apertures in either a flat or a corrugated metal film, providing a larger transmission of electromagnetic fields than would be expected from the small aperture size. Since its discovery in 1998, EOT has been a very active research field, leading both to the discovery of new ways of enhancing optical transmission and to its application to sensing, color filters, metamaterials, lenses, optical trapping, enhancement of nonlinear effects, among others. This paper reviews the different mechanisms that lead to EOT, paying special attention to the new research areas and applications that have appeared in the last few years.


Roadmap on structured light

Halina Rubinsztein-Dunlop, Andrew Forbes, M V Berry, M R Dennis, David L Andrews, Masud Mansuripur, Cornelia Denz, Christina Alpmann, Peter Banzer, Thomas Bauer, Ebrahim Karimi, Lorenzo Marrucci, Miles Padgett, Monika Ritsch-Marte, Natalia M Litchinitser, Nicholas P Bigelow, C Rosales-Guzmán, A Belmonte, J P Torres, Tyler W Neely, Mark Baker, Reuven Gordon, Alexander B Stilgoe, Jacquiline Romero, Andrew G White, Robert Fickler, Alan E Willner, Guodong Xie, Benjamin McMorran and Andrew M Weiner

Structured light refers to the generation and application of custom light fields. As the tools and technology to create and detect structured light have evolved, steadily the applications have begun to emerge. This roadmap touches on the key fields within structured light from the perspective of experts in those areas, providing insight into the current state and the challenges their respective fields face. Collectively the roadmap outlines the venerable nature of structured light research and the exciting prospects for the future that are yet to be realized.


Nonequilibrium dissipation in living oocytes

É. Fodor, W. W. Ahmed, M. Almonacid, M. Bussonnier, N. S. Gov, M.-H. Verlhac, T. Betz, P. Visco and F. van Wijland

Living organisms are inherently out-of-equilibrium systems. We employ recent developments in stochastic energetics and rely on a minimal microscopic model to predict the amount of mechanical energy dissipated by such dynamics. Our model includes complex rheological effects and nonequilibrium stochastic forces. By performing active microrheology and tracking micron-sized vesicles in the cytoplasm of living oocytes, we provide unprecedented measurements of the spectrum of dissipated energy. We show that our model is fully consistent with the experimental data, and we use it to offer predictions for the injection and dissipation energy scales involved in active fluctuations.


Wednesday, January 18, 2017

Optical dipole forces: Working together

Clarice D. Aiello

Strength lies in numbers and in teamwork: tens of thousands of artificial atoms tightly packed in a nanodiamond act cooperatively, enhancing the optical trapping forces beyond the expected classical bulk polarizability contribution.


Thermal fluctuation analysis of singly optically trapped spheres in hollow photonic crystal cavities

M. Tonin, F. M. Mor, L. Forró, S. Jeney, and R. Houdré

We report on the behaviour of singly optically trapped nanospheres inside a hollow, resonant photonic crystal cavity and measure experimentally the trapping constant using back-focal plane interferometry. We observe two trapping regimes arising from the back-action effect on the motion of the nanosphere in the optical cavity. The specific force profiles from these trapping regimes is measured.


Self-accelerating fan-shaped beams along arbitrary trajectories: a new tool for optical manipulation

Xiaolin Sui, Juanying Zhao, Bo Liu, Ziheng Yan, Changdong Cao and Shouhuan Zhou

We demonstrate, both theoretically and experimentally, a kind of fan-shaped optical beam propagating along the arbitrary trajectories (such as parabolic, hyperbolic and three-dimensional spiraling trajectories). With a controlled profile, this fan-shaped optical beam can be obtained from superposition of the Bessel-like beam and vortex Bessel-like beam. Also, the ability of guiding and transporting microparticles along its lobes is explored. These beams may find a variety of applications in optical trapping and manipulation.


Trapping and rotating of a metallic particle trimer with optical vortex

Z. Shen, L. Su, X.-C. Yuan, and Y.-C. Shen

We have experimentally observed the steady rotation of a mesoscopic size metallic particle trimer that is optically trapped by tightly focused circularly polarized optical vortex. Our theoretical analysis suggests that a large proportion of the radial scattering force pushes the metallic particles together, whilst the remaining portion provides the centripetal force necessary for the rotation. Furthermore, we have achieved the optical trapping and rotation of four dielectric particles with optical vortex. We found that, different from the metallic particles, instead of being pushed together by the radial scattering force, the dielectric particles are trapped just outside the maximum intensity ring of the focused field. The radial gradient force attracting the dielectric particles towards the maximum intensity ring provides the centripetal force for the rotation. The achieved steady rotation of the metallic particle trimer reported here may open up applications such as the micro-rotor.


Single-Molecule Force Spectroscopy Trajectories of a Single Protein and Its Polyproteins Are Equivalent: A Direct Experimental Validation Based on A Small Protein NuG2

Hai Lei, Dr. Chengzhi He, Prof. Dr. Chunguang Hu, Jinliang Li, Prof. Dr. Xiaodong Hu, Prof. Dr. Xiaotang Hu, Prof. Dr. Hongbin Li

Single-molecule force spectroscopy (SMFS) has become a powerful tool in investigating the mechanical unfolding/folding of proteins at the single-molecule level. Polyproteins made of tandem identical repeats have been widely used in atomic force microscopy (AFM)-based SMFS studies, where polyproteins not only serve as fingerprints to identify single-molecule stretching events, but may also improve statistics of data collection. However, the inherent assumption of such experiments is that all the domains in the polyprotein are equivalent and one SMFS trajectory of stretching a polyprotein made of n domains is equivalent to n trajectories of stretching a single domain. Such an assumption has not been validated experimentally. Using a small protein NuG2 and its polyprotein (NuG2)4 as model systems, here we use optical trapping (OT) to directly validate this assumption. Our results show that OT experiments on NuG2 and (NuG2)4 lead to identical parameters describing the unfolding and folding kinetics of NuG2, demonstrating that indeed stretching a polyprotein of NuG2 is equivalent to stretching single NuG2 in force spectroscopy experiments and thus validating the use of polyproteins in SMFS experiments.


Tuesday, January 17, 2017

Theory and practice of simulation of optical tweezers

Ann A.M. Bui, Alexander B. Stilgoe, Isaac C.D. Lenton, Lachlan J. Gibson, Anatolii V. Kashchuk, Shu Zhang, Halina Rubinsztein-Dunlop, Timo A. Nieminen

Computational modelling has made many useful contributions to the field of optical tweezers. One aspect in which it can be applied is the simulation of the dynamics of particles in optical tweezers. This can be useful for systems with many degrees of freedom, and for the simulation of experiments. While modelling of the optical force is a prerequisite for simulation of the motion of particles in optical traps, non-optical forces must also be included; the most important are usually Brownian motion and viscous drag. We discuss some applications and examples of such simulations. We review the theory and practical principles of simulation of optical tweezers, including the choice of method of calculation of optical force, numerical solution of the equations of motion of the particle, and finish with a discussion of a range of open problems.


Photocontrolled Supramolecular Assembling of Azobenzene-Based Biscalix[4]arenes upon Starting and Stopping Laser Trapping

Ken-ichi Yuyama, Lionel Marcelis, Pei-Mei Su, Wen-Sheng Chung, and Hiroshi Masuhara

Laser trapping in chemistry covers various studies ranging from single molecules, nanoparticles, and quantum dots to crystallization and liquid–liquid phase separation of amino acids. In this work, a supramolecular assembly of azobenzene-based biscalix[4]arene is generated in ethyl acetate using laser trapping; its nucleation and growth are elucidated. No trapping behavior was observed when a 1064 nm laser beam was focused inside of the solution; however, interesting assembling phenomena were induced when it was shined at the air/solution interface. A single disk having two layers was first prepared at the focal point of ∼1 μm and then expanded to the size of a few tens of micrometers, although no optical force was exerted outside of the focal volume. Upon switching the trapping laser off, needles were generated at the outer layer of the assembly, giving a stable sea urchin-like morphology to the generated assembly. At a 30–50% dilution of the initial solution in ethyl acetate, a mushroom-like morphology was also observed. Laser trapping-induced assembly of azobenzene-based biscalix[4]arene was quite different from the sharp-ellipsoidal aggregates obtained by the spontaneous evaporation of the solution. These trapping phenomena were specifically observed for biscalix[4]arene in the trans conformation of azo-benzene moiety but not for the cis-form, suggesting that the laser trapping of this azobenzene-based biscalix[4]arene is photocontrollable. Dynamics and mechanism of the supramolecular assembling are considered, referring to laser trapping-induced nucleation and liquid–liquid phase separation of amino acids.


Effects of cytoskeletal drugs on actin cortex elasticity

Yareni A. Ayala, Bruno Pontes, Barbara Hissa, Ana Carolina M. Monteiro, Marcos Farina, Vivaldo Moura-Neto, Nathan B. Viana, H. Moysés Nussenzveig

Mechanical properties of cells are known to be influenced by the actin cytoskeleton. In this article, the action of drugs that interact with the actin cortex is investigated by tether extraction and rheology experiments using optical tweezers. The influences of Blebbistatin, Cytochalasin D and Jasplakinolide on the cell mechanical properties are evaluated. The results, in contradiction to current views for Jasplakinolide, show that all three drugs and treatments destabilize the actin cytoskeleton, decreasing the cell membrane tension. The cell membrane bending modulus increased when the actin cytoskeleton was disorganized by Cytochalasin D. This effect was not observed for Blebbistatin and Jasplakinolide. All drugs decreased by two-fold the cell viscoelastic moduli, but only Cytochalasin D was able to alter the actin network into a more fluid-like structure. The results can be interpreted as the interplay between the actin network and the distribution of myosins as actin cross-linkers in the cytoskeleton. This information may contribute to a better understanding of how the membrane and cytoskeleton are involved in cell mechanical properties, underlining the role that each one plays in these properties.


Photokinetic analysis of the forces and torques exerted by optical tweezers carrying angular momentum

Aaron Yevick, Daniel J. Evans, David G. Grier

The theory of photokinetic effects expresses the forces and torques exerted by a beam of light in terms of experimentally accessible amplitude and phase profiles. We use this formalism to develop an intuitive explanation for the performance of optical tweezers operating in the Rayleigh regime, including effects arising from the influence of light’s angular momentum. First-order dipole contributions reveal how a focused beam can trap small objects, and what features limit the trap’s stability. The first-order force separates naturally into a conservative intensity-gradient term that forms a trap and a non-conservative solenoidal term that drives the system out of thermodynamic equilibrium. Neither term depends on the light’s polarization; light’s spin angular momentum plays no role at dipole order. Polarization-dependent effects, such as trap-strength anisotropy and spin-curl forces, are captured by the second-order dipole-interference contribution to the photokinetic force. The photokinetic expansion thus illuminates how light’s angular momentum can be harnessed for optical micromanipulation, even in the most basic optical traps.


Theoretical description of effective heat transfer between two viscously coupled beads

A. Bérut, A. Imparato, A. Petrosyan, and S. Ciliberto

We analytically study the role of nonconservative forces, namely viscous couplings, on the statistical properties of the energy flux between two Brownian particles kept at different temperatures. From the dynamical model describing the system, we identify an energy flow that satisfies a fluctuation theorem both in the stationary and in transient states. In particular, for the specific case of a linear nonconservative interaction, we derive an exact fluctuation theorem that holds for any measurement time in the transient regime, and which involves the energy flux alone. Moreover, in this regime the system presents an interesting asymmetry between the hot and cold particles. The theoretical predictions are in good agreement with the experimental results already presented in our previous article [Imparato et al., Phys. Rev. Lett. 116, 068301 (2016)], where we investigated the thermodynamic properties of two Brownian particles, trapped with optical tweezers, interacting through a dissipative hydrodynamic coupling.


Monday, January 16, 2017

Laser-driven gel microtool for single-cell manipulation based on temperature control with a photothermal conversion material

T. Hayakawa, M. Kikukawa, H. Maruyama, and F. Arai

We propose a laser-driven hybrid gel microtool for stable single-cell manipulation. The microtool is made of a microbead dyed with multiwalled carbon nanotubes (MWNT) and thermosensitive poly (N-isopropylacrylamide) gel coating. The gel adheres to cells at high temperatures but not at low temperatures. We can manipulate single cells without direct laser irradiation by adhering the cells to the gel on the microtool using the cell-adhesion property of the gel. The microtool is heated by trapping it with optical tweezers to make its surface cell-adhesive during the manipulation. Furthermore, we can control the optical heating property of the microtool by dyeing the microbeads with MWNT ink. The laser-heating-induced temperature increase of the microtool can be controlled from 4.2 °C to 23.5 °C by varying the concentration of MWNT ink. We succeeded in fabricating the proposed microtool and demonstrated single-cell transportation using the microtool without direct laser irradiation of the cell.


Mechanical unfolding kinetics of the SRV-1 gag-pro mRNA pseudoknot: possible implications for −1 ribosomal frameshifting stimulation

Zhensheng Zhong, Lixia Yang, Haiping Zhang, Jiahao Shi, J. Jeya Vandana, Do Thuy Uyen Ha Lam, René C. L. Olsthoorn, Lanyuan Lu & Gang Chen

Minus-one ribosomal frameshifting is a translational recoding mechanism widely utilized by many RNA viruses to generate accurate ratios of structural and catalytic proteins. An RNA pseudoknot structure located in the overlapping region of the gag and pro genes of Simian Retrovirus type 1 (SRV-1) stimulates frameshifting. However, the experimental characterization of SRV-1 pseudoknot (un)folding dynamics and the effect of the base triple formation is lacking. Here, we report the results of our single-molecule nanomanipulation using optical tweezers and theoretical simulation by steered molecular dynamics. Our results directly reveal that the energetic coupling between loop 2 and stem 1 via minor-groove base triple formation enhances the mechanical stability. The terminal base pair in stem 1 (directly in contact with a translating ribosome at the slippery site) also affects the mechanical stability of the pseudoknot. The −1 frameshifting efficiency is positively correlated with the cooperative one-step unfolding force and inversely correlated with the one-step mechanical unfolding rate at zero force. A significantly improved correlation was observed between −1 frameshifting efficiency and unfolding rate at forces of 15–35 pN, consistent with the fact that the ribosome is a force-generating molecular motor with helicase activity. No correlation was observed between thermal stability and −1 frameshifting efficiency.

Manipulation of cells with laser microbeam scissors and optical tweezers: a review

Karl Otto Greulich

The use of laser microbeams and optical tweezers in a wide field of biological applications from genomic to immunology is discussed. Microperforation is used to introduce a well-defined amount of molecules into cells for genetic engineering and optical imaging.
The microwelding of two cells induced by a laser microbeam combines their genetic outfit.
Microdissection allows specific regions of genomes to be isolated from a whole set of chromosomes. Handling the cells with optical tweezers supports investigation on the attack of immune systems against diseased or cancerous cells. With the help of laser microbeams, heart infarction can be simulated, and optical tweezers support studies on the heartbeat. Finally, laser microbeams are used to induce DNA damage in living cells for studies on cancer and ageing.


Static three-dimensional topological solitons in fluid chiral ferromagnets and colloids

Paul J. Ackerman & Ivan I. Smalyukh

Three-dimensional (3D) topological solitons are continuous but topologically nontrivial field configurations localized in 3D space and embedded in a uniform far-field background, that behave like particles and cannot be transformed to a uniform state through smooth deformations. Many topologically nontrivial 3D solitonic fields have been proposed. Yet, according to the Hobart–Derrick theorem, physical systems cannot host them, except for nonlinear theories with higher-order derivatives such as the Skyrme–Faddeev model. Experimental discovery of such solitons is hindered by the need for spatial imaging of the 3D fields, which is difficult in high-energy physics and cosmology. Here we experimentally realize and numerically model stationary topological solitons in a fluid chiral ferromagnet formed by colloidal dispersions of magnetic nanoplates. Such solitons have closed-loop preimages—3D regions with a single orientation of the magnetization field. We discuss localized structures with different linking of preimages quantified by topological Hopf invariants. The chirality is found to help in overcoming the constraints of the Hobart–Derrick theorem, like in two-dimensional ferromagnetic solitons, dubbed ‘baby skyrmions’. Our experimental platform may lead to solitonic condensed matter phases and technological applications.


Modified method for computing the optical force of the plasmonics nanoparticle from the Maxwell stress tensor

Dong Wang, Jun Song, Maozhen Xiong, Guangsheng Wang, Xiao Peng, and Junle Qu

By controlling the optical force, optical tweezers can manipulate many kinds of small particles without mechanical contact. In the theoretical analysis of the optical force, conventional methods are based on the integration of the Maxwell stress tensor over the outer surface of the particle, while the Maxwell stress tensor is determined by the electromagnetic field distribution around the particle itself. However, we find that this conventional method may not be appropriate in most situations, as two main issues arise, especially for plasmonics nanoparticles because of the metal involved. The first is the selection of the relative permittivity on the interface between the particle and the background medium, while the second is the use of the divergence theorem. Here, we present an improved and more correct technique to compute the optical force of optical tweezers on the plasmonics nanoparticle. The analysis of an Au-Ag core–shell nanostructure, conducted by adopting this revised method, shows that the negative force is located not only at the Fano resonance but also at longer wavelengths.


Friday, January 13, 2017

Optically assisted trapping with high-permittivity dielectric rings: Towards optical aerosol filtration

Rasoul Alaee, Muamer Kadic, Carsten Rockstuhl, and Ali Passian

Controlling the transport, trapping, and filtering of nanoparticles is important for many applications. By virtue of their weak response to gravity and their thermal motion, various physical mechanisms can be exploited for such operations on nanoparticles. However, the manipulation based on optical forces is potentially most appealing since it constitutes a highly deterministic approach. Plasmonic nanostructures have been suggested for this purpose, but they possess the disadvantages of locally generating heat and trapping the nanoparticles directly on the surface. Here, we propose the use of dielectric rings made of high permittivity materials for trapping nanoparticles. Thanks to their ability to strongly localize the field in space, nanoparticles can be trapped without contact. We use a semi-analytical method to study the ability of these rings to trap nanoparticles. The results are supported by full-wave simulations. Application of the trapping concept to nanoparticle filtration is suggested.


Interplay between optical, viscous, and elastic forces on an optically trapped Brownian particle immersed in a viscoelastic fluid

P. Domínguez-García, László Forró, and Sylvia Jeney

We provide a detailed study of the interplay between the different interactions which appear in the Brownian motion of a micronsized sphere immersed in a viscoelastic fluid measured with optical trapping interferometry. To explore a wide range of viscous, elastic, and optical forces, we analyze two different viscoelastic solutions at various concentrations, which provide a dynamic polymeric structure surrounding the Brownian sphere. Our experiments show that, depending on the fluid, optical forces, even if small, slightly modify the complex modulus at low frequencies. Based on our findings, we propose an alternative methodology to calibrate this kind of experimental set-up when non-Newtonian fluids are used. Understanding the influence of the optical potential is essential for a correct interpretation of the mechanical properties obtained by optically-trapped probe-based studies of biomaterials and living matter.


Role of the electromagnetic momentum in the spin-orbit interaction

Gianfranco Spavieri

The role played by the linear and angular momentum of the electromagnetic fields in the understanding of several aspects of quantum mechanics is discussed. A non-relativistic semi-classical model of the spin-orbit interaction, where the electromagnetic interaction energy U is calculated in the frame of the nucleus, is presented. Taking into account the electron hidden momentum Ph = c-1μ × E, the spin-orbit energy splitting turns out to be Δℰso = (1 / 2)U, the factor 1 / 2 emerging directly by requiring that the energy variation be a minimum. After quantization, the radius of the orbit is found to be spin-dependent, anticipating a feature of the Dirac equation. Finally, a test of the hidden momentum Ph, which may corroborate the approaches based on the hidden momentum and related interpretations of electrodynamics, is proposed and shown to be viable with present technology.


Surface-Plasmon-Mediated Gradient Force Enhancement and Mechanical State Transitions of Graphene Sheets

Peng Zhang, Nian-Hai Shen, Thomas Koschny, and Costas M. Soukoulis

Graphene, a two-dimensional material possessing extraordinary properties in electronics as well as mechanics, provides a great platform for various optoelectronic and optomechanical devices. Here, we theoretically study the optical gradient force arising from the coupling of surface plasmon modes on parallel graphene sheets, which can be several orders stronger than that between regular dielectric waveguides. Furthermore, with an energy functional optimization model, possible force-induced deformation of graphene sheets is calculated. We show that the significantly enhanced optical gradient force may lead to mechanical state transitions of graphene sheets, which are accompanied by abrupt changes in reflection and transmission spectra of the system. Our demonstrations illustrate the potential for broader graphene-related applications such as force sensors and actuators.


Lateral optical binding between two colloidal particles

Ming-Tzo Wei, Jack Ng, C. T. Chan & H. Daniel Ou-Yang

An optical binding force between two nearby colloidal particles trapped by two coherent laser beams is measured by phase-sensitive detection. The binding force is long-range and spatially oscillatory. For identical linearly-polarized incident beams, the oscillation period is equal to the optical wavelength. For mutually perpendicular polarizations, a new force appears with half-wavelength periodicity, caused by double inter-particle scattering. This force is observable only with cross-polarized incident beams, for which the stronger single-scattering forces are forbidden by parity.


Thursday, January 12, 2017

Study of in vitro RBCs membrane elasticity with AOD scanning optical tweezers

Huadong Song, Ying Liu, Bin Zhang, Kangzhen Tian, Panpan Zhu, Hao Lu, and Qi Tang

The elasticity of red cell membrane is a critical physiological index for the activity of RBC. Study of the inherent mechanism for RBCs membrane elasticity transformation is attention-getting all along. This paper proposes an optimized measurement method of erythrocytes membrane shear modulus incorporating acousto-optic deflector (AOD) scanning optical tweezers system. By use of this method, both membrane shear moduli and sizes of RBCs with different in vitro times were determined. The experimental results reveal that the RBCs membrane elasticity and size decline with in vitro time extension. In addition, semi quantitative measurements of S-nitrosothiol content in blood using fluorescent spectrometry during in vitro storage show that RBCs membrane elasticity change is positively associated with the S-nitrosylation level of blood. The analysis considered that the diminished activity of the nitric oxide synthase makes the S-nitrosylation of in vitro blood weaker gradually. The main reason for worse elasticity of the in vitro RBCs is that S-nitrosylation effect of spectrin fades. These results will provide a guideline for further study of in vitro cells activity and other clinical applications.


Orthogonal Nanoparticle Size, Polydispersity, and Stability Characterization with Near-Field Optical Trapping and Light Scattering

Perry Schein, Dakota O’Dell, and David Erickson

Here we present and demonstrate a new technique for simultaneously characterizing the size, polydispersity, and colloidal stability of nanoparticle suspensions. This method relies on tracking each nanoparticle’s motion in three spatial dimensions as it interacts with the evanescent field of an optical waveguide. The motion along the optical propagation axis of the waveguide provides insight into the polydispersity of a nanoparticle suspension. Horizontal motion perpendicular to the propagation axis gives the diffusion coefficient and particle size. In the direction normal to the surface, statistical analysis of the scattered light intensity distribution gives a map of the interaction energy landscape and insight into the suspension stability. These three orthogonal measurements are made simultaneously on each particle, building up population level insights from a single-particle rather than ensemble-averaged basis. We experimentally demonstrate the technique using polystyrene spheres obtaining results consistent with the manufacturer’s specifications for these suspensions. For NIST-traceable polystyrene size standard spheres, we measure a variability in the hydrodynamic radius of ±5 nm, compared with the manufacturer’s certified measurement of ±9 nm in the geometric diameter made using transmission electron microscopy.


Plasmonic structure: fiber grating formed by gold nanorods on a tapered fiber

J. O. Trevisanutto, A. Linhananta, and G. Das

The authors demonstrated the fabrication of a fiber Bragg grating-like plasmonic nanostructure on the surface of a tapered optical fiber using gold nanorods (GNRs). A multimode optical fiber with core and cladding diameters of 105 and 125 μm, respectively, was used to make a tapered fiber using a dynamic etching process. The tip diameter was ∼100 nm∼100 nm. Light from a laser was coupled to the untapered end of the fiber, which produced a strong evanescent field around the tapered section of the fiber. The gradient force due to the evanescent field trapped the GNRs on the surface of the tapered fiber. The authors explored possible causes of the GNR distribution. The plasmonic structure will be a good candidate for sensing based on surface enhanced Raman scattering.


Single-molecule analysis reveals multi-state folding of a guanine riboswitch

Vishnu Chandra, Zain Hannan, Huizhong Xu & Maumita Mandal

Guanine-responsive riboswitches undergo ligand-dependent structural rearrangements to control gene expression by transcription termination. While the molecular basis for ligand recognition is well established, the associated structural rearrangements and the kinetics involved in the formation of the aptamer domain are less well understood. Using high-resolution optical tweezers, we followed the folding trajectories of a single molecule of the xpt–pbuX guanine aptamer from Bacillus subtilis. We report a rapid six-state conformational rearrangement, in which three of the states are guanine dependent, during the transition from the linear to the native receptor conformation. The folding completes in <1 s. The force-dependent equilibrium kinetics and the mutational data indicated that the flexible J2–J3 junction undergoes a ligand-dependent conformational switching, which triggers the formation of the long-range tertiary interactions and the P1 helix. In the absence of the right ligand, the junction failed to initiate the series of conformational rearrangements required for the riboswitch activities.


A programmable DNA origami nanospring that reveals force-induced adjacent binding of myosin VI heads

M. Iwaki, S. F. Wickham, K. Ikezaki, T. Yanagida & W. M. Shih

Mechanosensitive biological nanomachines such as motor proteins and ion channels regulate diverse cellular behaviour. Combined optical trapping with single-molecule fluorescence imaging provides a powerful methodology to clearly characterize the mechanoresponse, structural dynamics and stability of such nanomachines. However, this system requires complicated experimental geometry, preparation and optics, and is limited by low data-acquisition efficiency. Here we develop a programmable DNA origami nanospring that overcomes these issues. We apply our nanospring to human myosin VI, a mechanosensory motor protein, and demonstrate nanometre-precision single-molecule fluorescence imaging of the individual motor domains (heads) under force. We observe force-induced transitions of myosin VI heads from non-adjacent to adjacent binding, which correspond to adapted roles for low-load and high-load transport, respectively. Our technique extends single-molecule studies under force and clarifies the effect of force on biological processes.


Wednesday, January 11, 2017

Plasmonic Heating-Assisted Laser-Induced Crystallization from a NaClO3 Unsaturated Mother Solution

Hiromasa Niinomi, Teruki Sugiyama, Miho Tagawa, Mihoko Maruyama, Toru Ujihara, Takashige Omatsu, and Yusuke Mori

We provide a novel laser-induced crystallization mechanism which explains crystallization induced by visible laser trapping of silver nanoparticles (AgNPs) at the air/unsaturated mother solution interface from the focal spot [Niinomi et al.CrystEngComm 2016, 18, 7441–7448]. Simultaneous in situ microscopic observation of Raman scattering and polarized-light image revealed that the optical trapping of nanoparticles that exhibit surface-enhanced Raman scattering (SERS) triggers the crystallization, showing the excitation of localized surface plasmon resonance (LSPR) significantly promotes the crystallization. Numerical analysis of temperature distribution based on the combination of finite-difference time-domain electromagnetic and finite-difference heat transfer calculations shows that temperature reaches 390 °C at the focal spot because of plasmonic heating, the energy dissipation of the plasmon-enhanced electromagnetic field as heat. A conceivable mechanism of the crystallization is local increment of supersaturation caused by local solvent evaporation via the Plasmonic heating. This plasmonic heating assisted laser-induced nucleation process has the possibility to provide not only a novel approach for spatiotemporal control of crystallization but also a novel nucleation field based on nonlinear light–matter interaction originating from the plasmon-enhanced electromagnetic near field through heterogeneous nucleation on the surface of plasmonic particles.


Highly Sensitive Detection of Organic Molecules on the Basis of a Poly(N-isopropylacrylamide) Microassembly Formed by Plasmonic Optical Trapping

Tatsuya Shoji, Daiki Sugo, Fumika Nagasawa, Kei Murakoshi, Noboru Kitamura, and Yasuyuki Tsuboi

We demonstrate that a poly(N-isopropylacrylamide) (PNIPAM) microassembly, formed by plasmonic optical trapping, can provide the platform for a highly sensitive detection technique for fluorescent and nonfluorescent organic molecules dissolved in aqueous solution. PNIPAM microassemblies can be easily formed by a combination with a photothermal effect and an enhanced optical force. These physical phenomena were obtained through resonant excitation of localized surface plasmon (LSP). Sparsely distributed fluorescent or nonfluorescent molecules dissolved in solution can be extracted into the PNIPAM assembly, resulting in an increase in fluorescence or Raman signals. In particular, we successfully detected quite small amounts of analytes (rhodamine B) at the 10–9 mol/L level. Using LSP is an alternative approach in analytical chemistry and can be used in addition to surface enhanced Raman scattering and surface enhanced fluorescence.


Early-Onset Hypertrophic Cardiomyopathy Mutations Significantly Increase the Velocity, Force, and Actin-Activated ATPase Activity of Human β-Cardiac Myosin

Arjun S. Adhikari, Kristina B. Kooiker, Saswata S. Sarkar, Chao Liu, Daniel Bernstein, James A. Spudich, Kathleen M. Ruppel

Hypertrophic cardiomyopathy (HCM) is a heritable cardiovascular disorder that affects 1 in 500 people. A significant percentage of HCM is attributed to mutations in β-cardiac myosin, the motor protein that powers ventricular contraction. This study reports how two early-onset HCM mutations, D239N and H251N, affect the molecular biomechanics of human β-cardiac myosin. We observed significant increases (20%–90%) in actin gliding velocity, intrinsic force, and ATPase activity in comparison to wild-type myosin. Moreover, for H251N, we found significantly lower binding affinity between the S1 and S2 domains of myosin, suggesting that this mutation may further increase hyper-contractility by releasing active motors. Unlike previous HCM mutations studied at the molecular level using human β-cardiac myosin, early-onset HCM mutations lead to significantly larger changes in the fundamental biomechanical parameters and show clear hyper-contractility.


Long-range interactions, wobbles, and phase defects in chains of model cilia

Douglas R. Brumley, Nicolas Bruot, Jurij Kotar, Raymond E. Goldstein, Pietro Cicuta, and Marco Polin

Eukaryotic cilia and flagella are chemo-mechanical oscillators capable of generating long-range coordinated motions known as metachronal waves. Pair synchronization is a fundamental requirement for these collective dynamics, but it is generally not sufficient for collective phase-locking, chiefly due to the effect of long-range interactions. Here we explore experimentally and numerically a minimal model for a ciliated surface: hydrodynamically coupled oscillators rotating above a no-slip plane. Increasing their distance from the wall profoundly affects the global dynamics, due to variations in hydrodynamic interaction range. The array undergoes a transition from a traveling wave to either a steady chevron pattern or one punctuated by periodic phase defects. Within the transition between these regimes the system displays behavior reminiscent of chimera states.


Poly(3-hydroxybutyrate) anabolism in Cupriavidus necator cultivated at various carbon-to-nitrogen ratios: insights from single-cell Raman spectroscopy

Zhanhua Tao ; Pengfei Zhang ; Zhaojun Qin ; Yong-Qing Li ; Guiwen Wang

Cupriavidus necator accumulates large amounts of poly(3-hydroxybutyrate) (PHB), a biodegradable substitute for petroleum-based plastics, under certain nutrient conditions. Conventional solvent-extraction-based methods for PHB quantification only obtain average information from cell populations and, thus, mask the heterogeneity among individual cells. Laser tweezers Raman spectroscopy (LTRS) was used to monitor dynamic changes in the contents of PHB, nucleic acids, and proteins in C. necator at the population and single-cell levels when the microorganism cells were cultivated at various carbon-to-nitrogen ratios. The biosynthetic activities of nucleic acids and proteins were maintained at high levels, and only a small amount of PHB was produced when the bacterial cells were cultured under balanced growth conditions. By contrast, the syntheses of nucleic acids and proteins were blocked, and PHB was accumulated in massive amount inside the microbial cells under nitrogen-limiting growth circumstances. Single-cell analysis revealed a relatively high heterogeneity in PHB level at the early stage of the bacterial growth. Additionally, bacterial cells in populations at certain cultivation stages were composed of two or three subpopulations on the basis of their PHB abundance. Overall, LTRS is a reliable single-cell analysis tool that can provide insights into PHB fermentation.


Tuesday, January 10, 2017

Nanoscopy of bacterial cells immobilized by holographic optical tweezers

Robin Diekmann, Deanna L. Wolfson, Christoph Spahn, Mike Heilemann, Mark Schüttpelz & Thomas Huser

Imaging non-adherent cells by super-resolution far-field fluorescence microscopy is currently not possible because of their rapid movement while in suspension. Holographic optical tweezers (HOTs) enable the ability to freely control the number and position of optical traps, thus facilitating the unrestricted manipulation of cells in a volume around the focal plane. Here we show that immobilizing non-adherent cells by optical tweezers is sufficient to achieve optical resolution well below the diffraction limit using localization microscopy. Individual cells can be oriented arbitrarily but preferably either horizontally or vertically relative to the microscope’s image plane, enabling access to sample sections that are impossible to achieve with conventional sample preparation and immobilization. This opens up new opportunities to super-resolve the nanoscale organization of chromosomal DNA in individual bacterial cells.


Intact Telopeptides Enhance Interactions between Collagens

Marjan Shayegan, Tuba Altindal, Evan Kiefl, Nancy R. Forde

Collagen is the fundamental structural component of a wide range of connective tissues and of the extracellular matrix. It undergoes self-assembly from individual triple-helical proteins into well-ordered fibrils, a process that is key to tissue development and homeostasis, and to processes such as wound healing. Nucleation of this assembly is known to be slowed considerably by pepsin removal of short nonhelical regions that flank collagen’s triple helix, known as telopeptides. Using optical tweezers to perform microrheology measurements, we explored the changes in viscoelasticity of solutions of collagen with and without intact telopeptides. Our experiments reveal that intact telopeptides contribute a significant frequency-dependent enhancement of the complex shear modulus. An analytical model of polymers associating to establish chemical equilibrium among higher-order species shows trends in G′ and G″ consistent with our experimental observations, including a concentration-dependent crossover in G″/c around 300 Hz. This work suggests that telopeptides facilitate transient intermolecular interactions between collagen proteins, even in the acidic conditions used here.


Statistical Thermodynamic Model for Surface Tension of Organic and Inorganic Aqueous Mixtures

Hallie C. Boyer, Bryan R. Bzdek, Jonathan P. Reid, and Cari S. Dutcher

The surface composition and tensions of aqueous aerosols govern a set of processes that largely determine the fate of particles in the atmosphere. Predictive modeling of surface tension can provide significant contributions to studies of atmospheric aerosol effects on climate and human health. A previously derived surface tension model for single solute aqueous solutions used adsorption isotherms and statistical mechanics to enable surface tension predictions across the entire concentration range as a function of solute activity. Here, we extend the model derivation to address multicomponent solutions and demonstrate its accuracy with systems containing mixtures of electrolytes and organic solutes. Binary model parameters are applied to the multicomponent model, requiring no further parametrization for mixtures. Five ternary systems are studied here and represent three types of solute combinations: organic–organic (glycerol–ethanol), electrolyte–organic (NaCl–succinic acid, NaCl–glutaric acid), and electrolyte–electrolyte (NaCl–KCl and NH4NO3–(NH4)2SO4). For the NaCl-glutaric acid system, experimental measurements of picoliter droplet surface tension using aerosol optical tweezers show excellent agreement with the model predictions.


Active bioparticle manipulation in microfluidic systems

Mohd Anuar Md Ali, Kostya (Ken) Ostrikov, Fararishah Abdul Khalid, Burhanuddin Y. Majlis and Aminuddin A. Kayani

The motion of bioparticles in a microfluidic environment can be actively controlled using several tuneable mechanisms, including hydrodynamic, electrophoresis, dielectrophoresis, magnetophoresis, acoustophoresis, thermophoresis and optical forces. These mechanisms are applied to obtain desired bioparticle motions which are important in facilitating different biological processes. In this work, we review the fundamentals, features and applications of these tuneable mechanisms for the manipulation of bioparticles such as proteins, nucleic acids, viruses, bacteria, stem cells, cancer and tumor cells, blood cells and multicellular organisms in microfluidic systems. We focus on applications that can realize biomedical devices potentially suitable in diagnostic, therapeutic or analytical applications. Future perspectives of microfluidic systems incorporating active bioparticle manipulation mechanisms are included.


Real-Time, Label-Free Detection of Local Exocytosis Outside Pancreatic β Cells Using Laser Tweezers Raman Spectroscopy

Rui-qiong Luo, Fang Wei, Shu-shi Huang, Yue-ming Jiang, Shan-lei Zhang, Wen-qing Mo, Hong Liu, Xi Rong

The examination of insulin (Ins) exocytosis at the single-cell level by conventional methods, such as electrophysiological approaches, total internal reflection imaging, and two-photon imaging technology, often requires an invasive microelectrode puncture or label. In this study, high concentrations of glucose and potassium chloride were used to stimulate β cell Ins exocytosis, while low concentrations of glucose and calcium channel blockers served as the blank and negative control, respectively. Laser tweezers Raman spectroscopy (LTRS) was used to capture the possible Raman scattering signal from a local zone outside of the cell edge. The results show that the frequencies of the strong signals from the local zones outside the cellular edge in the stimulated groups are greater than those of the control. The Raman spectra from the cellular edge, Ins and cell membrane were compared. Thus, local Ins exocytosis activity outside pancreatic β cells might be observed indirectly using LTRS, a non-invasive optical method.


Monday, January 9, 2017

Behaviors of ellipsoidal micro-particles within a two-beam optical levitator

T. Petkov, M. Yang, K.F. Ren, B. Pouligny, J.-C. Loudet

The two-beam levitator (TBL) is a standard optical setup made of a couple of counter-propagating beams. Note worthily, TBLs allow the manipulation and trapping of particles at long working distances. While much experience has been accumulated in the trapping of single spherical particles in TBLs, the behaviors of asymmetrical particles turn out to be more complex, and even surprising. Here, we report observations with prolate ellipsoidal polystyrene particles, with varying aspect ratio and ratio of the two beam powers. Generalizing the earlier work by Mihiretie et al. in single beam geometries [JQSRT 126, 61 (2013)], we observe that particles may be either static, or permanently oscillating, and that the two-beam geometry produces new particle responses: some of them are static, but non-symmetrical, while others correspond to new types of oscillations. A two-dimensional model based on ray-optics qualitatively accounts for these configurations and for the “primary” oscillations of the particles. Furthermore, levitation powers measured in the experiments are in fair agreement with those computed from GLMT (Generalized Lorentz Mie Theory), MLFMA (Multilevel Fast Multipole Algorithm) and approximate ray-optics methods.


Routing light with ultrathin nanostructures beyond the diffraction limit

Haiyang Huang, Aimin Wu, Hao Li, Wei Li, Zhen Sheng, Shichang Zou, Xi Wang, and Fuwan Gan

An open nanostructure consisting of a periodic chain of subwavelength-nanoparticles for compressing and routing light beyond the diffraction limit is proposed. The open nanostructure is ultrathin and compact, with a size much smaller than the wavelength of light. We demonstrate that our ultrathin open nanostructure provides functions that can route and manipulate light at the subwavelength scale and can also sharply bend and split light beams below the diffraction limit while exhibiting broadband, incident-angle-tolerant, and robust against disorder. A physical picture based on all-angle self-collimation is presented to understand the manipulation of light using the ultrathin open nanostructure. Experimental and numerical observations validate our findings. This approach provides great flexibility in the design of nanophotonic devices for routing and manipulating light beyond the diffraction limit.


Transverse optical forces for manipulating nanoparticles

Alexander A. Zharov, Alexander A. Zharov, Jr., Ilya V. Shadrivov, and Nina A. Zharova

We study optical forces acting on a subwavelength particle with anisotropic polarizability and discover an optomechanical effect that resembles the Hall effect for electrons. While in the classical Hall effect the transverse Lorentz force and the transverse voltage appear due to the static magnetic field which induces the nondiagonal components of the electric conductivity tensor; in our case the imaginary parts of the nondiagonal elements of the polarizability tensor are responsible for the transverse scattering force. We calculate this force for the examples of the ellipsoidal plasmonic nanoparticles and the spherical particle with gyromagnetic properties, and show that the transverse force depends on the physical origin of the anisotropy of the polarizability, and on the electromagnetic wave structure around the particle. Moreover, this force primarily occurs in the inhomogeneous field only.


Submillimetre Network Formation by Light-induced Hybridization of Zeptomole-level DNA

Takuya Iida, Yushi Nishimura, Mamoru Tamura, Keisuke Nishida, Syoji Ito & Shiho Tokonami

Macroscopic unique self-assembled structures are produced via double-stranded DNA formation (hybridization) as a specific binding essential in biological systems. However, a large amount of complementary DNA molecules are usually required to form an optically observable structure via natural hybridization, and the detection of small amounts of DNA less than femtomole requires complex and time-consuming procedures. Here, we demonstrate the laser-induced acceleration of hybridization between zeptomole-level DNA and DNA-modified nanoparticles (NPs), resulting in the assembly of a submillimetre network-like structure at the desired position with a dramatic spectral modulation within several minutes. The gradual enhancement of light-induced force and convection facilitated the two-dimensional network growth near the air-liquid interface with optical and fluidic symmetry breakdown. The simultaneous microscope observation and local spectroscopy revealed that the assembling process and spectral change are sensitive to the DNA sequence. Our findings establish innovative guiding principles for facile bottom-up production via various biomolecular recognition events.


DNA intercalation optimized by two-step molecular lock mechanism

Ali A. Almaqwashi, Johanna Andersson, Per Lincoln, Ioulia Rouzina, Fredrik Westerlund & Mark C. Williams

The diverse properties of DNA intercalators, varying in affinity and kinetics over several orders of magnitude, provide a wide range of applications for DNA-ligand assemblies. Unconventional intercalation mechanisms may exhibit high affinity and slow kinetics, properties desired for potential therapeutics. We used single-molecule force spectroscopy to probe the free energy landscape for an unconventional intercalator that binds DNA through a novel two-step mechanism in which the intermediate and final states bind DNA through the same mono-intercalating moiety. During this process, DNA undergoes significant structural rearrangements, first lengthening before relaxing to a shorter DNA-ligand complex in the intermediate state to form a molecular lock. To reach the final bound state, the molecular length must increase again as the ligand threads between disrupted DNA base pairs. This unusual binding mechanism results in an unprecedented optimized combination of high DNA binding affinity and slow kinetics, suggesting a new paradigm for rational design of DNA intercalators.