Monday, September 18, 2017

Accuracy and Mechanistic Details of Optical Printing of Single Au and Ag Nanoparticles

Julián Gargiulo, Ianina L. Violi, Santiago Cerrota, Lukáš Chvátal, Emiliano Cortés, Eduardo M. Perassi, Fernando Diaz, Pavel Zemánek, and Fernando D. Stefani

Optical printing is a powerful all-optical method that allows the incorporation of colloidal nanoparticles (NPs) onto substrates with nanometric precision. Here, we present a systematic study of the accuracy of optical printing of Au and Ag NPs, using different laser powers and wavelengths. When using light of wavelength tuned to the localized surface plasmon resonance (LSPR) of the NPs, the accuracy improves as the laser power is reduced, whereas for wavelengths off the LSPR, the accuracy is independent of the laser power. Complementary studies of the printing times of the NPs reveal the roles of Brownian and deterministic motion. Calculated trajectories of the NPs, taking into account the interplay between optical forces, electrostatic forces, and Brownian motion, allowed us to rationalize the experimental results and gain a detailed insight into the mechanism of the printing process. A clear framework is laid out for future optimizations of optical printing and optical manipulation of NPs near substrates.


Rigorous full-wave calculation of optical forces on dielectric and metallic microparticles immersed in a vector Airy beam

Wanli Lu, Huajin Chen, Shiyang Liu, and Zhifang Lin

Based on the generalized Lorenz-Mie theory and the Maxwell stress tensor approach we present the first rigorous full-wave solution of the optical forces acting on spherical microparticles immersed in a two-dimensional vector Airy beam beyond the paraxial approximation. The critical aspect lies in evaluating efficiently and accurately the partial wave expansion coefficients of the incident Airy beam, which are achieved by using the vector angular spectrum representation for a variety of polarizations. The optical field distributions are then simulated to show the self-accelerating and self-healing effects of the Airy beam. The dielectric and gold microparticles are shown to be trapped within the main lobe or the nearby side-lobes mostly by the transverse gradient optical force while driven forward along the parabolic trajectory of the Airy beam by the longitudinal scattering force. It is thus demonstrated theoretically that the vector Airy beam has the capability of precisely transporting both dielectric and metallic microparticles along the prespecified curved paths.


Mechanical measurement of hydrogen bonded host–guest systems under non-equilibrium, near-physiological conditions

Teresa Naranjo, Fernando Cerrón, Belén Nieto-Ortega, Alfonso Latorre, Álvaro Somoza, Borja Ibarra and Emilio M. Pérez

Decades after the birth of supramolecular chemistry, there are many techniques to measure noncovalent interactions, such as hydrogen bonding, under equilibrium conditions. As ensembles of molecules rapidly lose coherence, we cannot extrapolate bulk data to single-molecule events under non-equilibrium conditions, more relevant to the dynamics of biological systems. We present a new method that exploits the high force resolution of optical tweezers to measure at the single molecule level the mechanical strength of a hydrogen bonded host–guest pair out of equilibrium and under near-physiological conditions. We utilize a DNA reporter to unambiguously isolate single binding events. The Hamilton receptor–cyanuric acid host–guest system is used as a test bed. The force required to dissociate the host–guest system is ∼17 pN and increases with the pulling rate as expected for a system under non-equilibrium conditions. Blocking one of the hydrogen bonding sites results in a significant decrease of the force-to-break by 1–2 pN, pointing out the ability of the method to resolve subtle changes in the mechanical strength of the binding due to the individual H-bonding components. We believe the method will prove to be a versatile tool to address important questions in supramolecular chemistry.


Force spectroscopy unravels the role of ionic strength on DNA-cisplatin interaction: Modulating the binding parameters

L. Oliveira and M. S. Rocha

In the present work we have gone a step forward in the understanding of the DNA-cisplatin interaction, investigating the role of the ionic strength on the complexes formation. To achieve this task, we use optical tweezers to perform force spectroscopy on the DNA-cisplatin complexes, determining their mechanical parameters as a function of the drug concentration in the sample for three different buffers. From such measurements, we determine the binding parameters and study their behavior as a function of the ionic strength. The equilibrium binding constant decreases with the counterion concentration ([Na]) and can be used to estimate the effective net charge of cisplatin in solution. The cooperativity degree of the binding reaction, on the other hand, increases with the ionic strength, as a result of the different conformational changes induced by the drug on the double-helix when binding under different buffer conditions. Such results can be used to modulate the drug binding to DNA, by appropriately setting the ionic strength of the surrounding buffer. The conclusions drawn provide significant new insights on the complex cooperative interactions between the DNA molecule and the class of platinum-based compounds, much used in chemotherapies.


Membrane Tension: A Challenging But Universal Physical Parameter in Cell Biology

BrunoPontes, Pascale Monzo Nils C.Gauthier

The plasma membrane separates the interior of cells from the outside environment. The membrane tension, defined as the force per unit length acting on a cross-section of membrane, regulates many vital biological processes. In this review, we summarize the first historical findings and the latest advances, showing membrane tension as an important physical parameter in cell biology. We also discuss how this parameter must be better integrated and we propose experimental approaches for key unanswered questions.


Thursday, September 14, 2017

Force-activated DNA substrates for probing individual proteins interacting with single-stranded DNA

Stephen R. Okoniewski Lyle Uyetake Thomas T. Perkins

Single-molecule force spectroscopy provides insight into how proteins bind to and move along DNA. Such studies often embed a single-stranded (ss) DNA region within a longer double-stranded (ds) DNA molecule. Yet, producing these substrates remains laborious and inefficient, particularly when using the traditional three-way hybridization. Here, we developed a force-activated substrate that yields an internal 1000 nucleotide (nt) ssDNA region when pulled partially into the overstretching transition (∼65 pN) by engineering a 50%-GC segment to have no adjacent GC base pairs. Once the template was made, these substrates were efficiently prepared by polymerase chain reaction amplification followed by site-specific nicking. We also generated a more complex structure used in high-resolution helicase studies, a DNA hairpin adjacent to 33 nt of ssDNA. The temporally defined generation of individual hairpin substrates in the presence of RecQ helicase and saturating adenine triphosphate let us deduce that RecQ binds to ssDNA via a near diffusion-limited reaction. More broadly, these substrates enable the precise initiation of an important class of protein–DNA interactions.


Z-ring Structure and Constriction Dynamics in E. coli

Pramod Kumar, Amarjeet Yadav, Itzhak Fishov and Mario Feingold

The Z-ring plays a central role in bacterial division. It consists of FtsZ filaments, but the way these reorganize in the ring-like structure during septation remains largely unknown. Here, we measure the effective constriction dynamics of the ring. Using an oscillating optical trap, we can switch individual rod-shaped E. coli cells between horizontal and vertical orientations. In the vertical orientation, the fluorescent Z-ring image appears as a symmetric circular structure that renders itself to quantitative analysis. In the horizontal orientation, we use phase-contrast imaging to determine the extent of the cell constriction and obtain the effective time of division. We find evidence that the Z-ring constricts at a faster rate than the cell envelope such that its radial width (inwards from the cytoplasmic membrane) grows during septation. In this respect, our results differ from those recently obtained using photoactivated localization microscopy (PALM) where the radial width of the Z-ring was found to be approximately constant as the ring constricts. A possible reason for the different behavior of the constricting Z-rings could be the significant difference in the corresponding cell growth rates.


Accurate nanoscale flexibility measurement of DNA and DNA–protein complexes by atomic force microscopy in liquid

Divakaran Murugesapillai, Serge Bouaziz, L. James Maher, III, Nathan E. Israeloff, Craig E. Cameron and Mark C. Williams

The elasticity of double-stranded DNA (dsDNA), as described by its persistence length, is critical for many biological processes, including genomic regulation. A persistence length value can be obtained using atomic force microscopy (AFM) imaging. However, most AFM studies have been done by depositing the sample on a surface using adhesive ligands and fitting the contour to a two-dimensional (2D) wormlike chain (WLC) model. This often results in a persistence length measurement that is different from the value determined using bulk and single molecule methods. We describe a method for obtaining accurate three-dimensional (3D) persistence length measurements for DNA and DNA–protein complexes by using a previously developed liquid AFM imaging method and then applying the 3D WLC model. To demonstrate the method, we image in both air and liquid several different dsDNA constructs and DNA–protein complexes that both increase (HIV-1 Vpr) and decrease (yeast HMO1) dsDNA persistence length. Fitting the liquid AFM-imaging contour to the 3D WLC model results in a value in agreement with measurements obtained in optical tweezers experiments. Because AFM also allows characterization of local DNA properties, the ability to correctly measure global flexibility will strongly increase the impact of measurements that use AFM imaging.


Parametric feedback cooling of levitated optomechanics in a parabolic mirror trap

Jamie Vovrosh, Muddassar Rashid, David Hempston, James Bateman, Mauro Paternostro, and Hendrik Ulbricht

Levitated optomechanics, a new experimental physics platform, holds promise for fundamental science and quantum technological sensing applications. We demonstrate a simple and robust geometry for optical trapping in vacuum of a single nanoparticle based on a parabolic mirror and the optical gradient force. We demonstrate parametric feedback cooling of all three motional degrees of freedom from room temperature to a few millikelvin. A single laser at 1550 nm and a single photodiode are used for trapping, position detection, and cooling for all three dimensions. Particles with diameters from 26 to 160 nm are trapped without feedback to 10−5 mbar10−5 mbar, and with feedback-engaged, the pressure is reduced to 10−6 mbar10−6 mbar. Modifications to the harmonic motion in the presence of noise and feedback are studied, and an experimental mechanical quality factor in excess of 4×1074×107 is evaluated. This particle manipulation is key to building a nanoparticle matter-wave interferometer in order to test the quantum superposition principle in the macroscopic domain.


Localized plasmonic structured illumination microscopy with optically trapped microlens

Anna Bezryadina, Jinxing Li, Junxiang Zhao, Alefia Kothambawala, Joseph Louis Ponsetto, Eric Huang, Joseph Wang and Zhaowei Liu

Localized plasmonic structured illumination microscopy (LPSIM) is a recently developed super resolution technique that demonstrates immense potential via arrays of localized plasmonic antennas. Microlens microscopy represents another distinct approach for improving resolution by introducing spherical lens with large refractive index to boost the effective numerical aperture of the imaging system. In this LetterPaper, we bridge together the LPSIM and optically trapped spherical microlenses, for the first time, to demonstrate a new super resolution technique for surface imaging. By trapping and moving polystyrene and TiO2 microspheres with optical tweezers on top of a LPSIM substrate, the new imaging system has achieved higher NA and resolution improvement.


Wednesday, September 13, 2017

Effect of red light on optically trapped spermatozoa

Kay W. Chow, Daryl Preece, and Michael W. Berns

Successful artificial insemination relies on the use of high quality spermatozoa. One measure of sperm quality is swimming force. Increased swimming force has been correlated with higher sperm swimming speeds and improved reproductive success. It is hypothesized that by increasing sperm swimming speed, one can increase swimming force. Previous studies have shown that red light irradiation causes an increase in sperm swimming speed. In the current study, 633nm red light irradiation is shown to increase mean squared displacement in trapped sperm. The methodology allows for comparison of relative swimming forces between irradiated and non-irradiated samples.


Roadmap for optofluidics

Paolo Minzioni, Roberto Osellame, Cinzia Sada, S Zhao, F G Omenetto, Kristinn B Gylfason, Tommy Haraldsson, Yibo Zhang, Aydogan Ozcan, Adam Wax

Optofluidics, nominally the research area where optics and fluidics merge, is a relatively new research field and it is only in the last decade that there has been a large increase in the number of optofluidic applications, as well as in the number of research groups, devoted to the topic. Nowadays optofluidics applications include, without being limited to, lab-on-a-chip devices, fluid-based and controlled lenses, optical sensors for fluids and for suspended particles, biosensors, imaging tools, etc. The long list of potential optofluidics applications, which have been recently demonstrated, suggests that optofluidic technologies will become more and more common in everyday life in the future, causing a significant impact on many aspects of our society. A characteristic of this research field, deriving from both its interdisciplinary origin and applications, is that in order to develop suitable solutions a combination of a deep knowledge in different fields, ranging from materials science to photonics, from microfluidics to molecular biology and biophysics, is often required. As a direct consequence, also being able to understand the long-term evolution of optofluidics research is not easy. In this article, we report several expert contributions on different topics so as to provide guidance for young scientists. At the same time, we hope that this document will also prove useful for funding institutions and stakeholders to better understand the perspectives and opportunities offered by this research field.


Size- and speed-dependent mechanical behavior in living mammalian cytoplasm

Jiliang Hu, Somaye Jafari, Yulong Han, Alan J. Grodzinsky, Shengqiang Cai, and Ming Guo

Active transport in the cytoplasm plays critical roles in living cell physiology. However, the mechanical resistance that intracellular compartments experience, which is governed by the cytoplasmic material property, remains elusive, especially its dependence on size and speed. Here we use optical tweezers to drag a bead in the cytoplasm and directly probe the mechanical resistance with varying size a and speed V. We introduce a method, combining the direct measurement and a simple scaling analysis, to reveal different origins of the size- and speed-dependent resistance in living mammalian cytoplasm. We show that the cytoplasm exhibits size-independent viscoelasticity as long as the effective strain rate V/a is maintained in a relatively low range (0.1 s−1 < V/a < 2 s−1) and exhibits size-dependent poroelasticity at a high effective strain rate regime (5 s−1 < V/a < 80 s−1). Moreover, the cytoplasmic modulus is found to be positively correlated with only V/a in the viscoelastic regime but also increases with the bead size at a constant V/a in the poroelastic regime. Based on our measurements, we obtain a full-scale state diagram of the living mammalian cytoplasm, which shows that the cytoplasm changes from a viscous fluid to an elastic solid, as well as from compressible material to incompressible material, with increases in the values of two dimensionless parameters, respectively. This state diagram is useful to understand the underlying mechanical nature of the cytoplasm in a variety of cellular processes over a broad range of speed and size scales.


Kinetically Determined Hygroscopicity and Efflorescence of Sucrose–Ammonium Sulfate Aerosol Droplets under Lower Relative Humidity

Lin-Na Wang, Chen Cai, and Yun-Hong Zhang

Organic aerosols will likely form in semisolid, glassy, and high viscous state in the atmosphere, which show nonequilibrium kinetic characteristics at low relative humidity (RH) conditions. In this study, we applied optical tweezers to investigate the water transport in a sucrose/(NH4)2SO4 droplet with high organic to inorganic mole ratio (OIR). The characteristic time ratio between the droplet radius and the RH was used to describe the water mass transfer difference dependent on RH. For OIR greater than 1:1 in sucrose/(NH4)2SO4 droplets, the characteristic time ratio at low RH (<∼30% RH) was two orders magnitude greater than that at high RH (>∼60%). We also coupled vacuum FTIR spectrometer and a high-speed photography to study the efflorescence process in sucrose/(NH4)2SO4 droplets with low OIR. The crystalline fraction of (NH4)2SO4 was used to understand efflorescence behavior when the RH was linearly decreasing with a velocity of 1.2% RH min–1. Because of suppression of (NH4)2SO4 nucleation by addition of sucrose, the efflorescence relative humidity (ERH) of (NH4)2SO4 decrease from the range of ∼48.2% to ∼36.1% for pure (NH4)2SO4 droplets to from ∼44.7% to ∼25.4%, from ∼43.2% to ∼21.2%, and from ∼41.7% to ∼21.1% for the mixed droplets with OIR of 1:4, 1:3, and 1:2, respectively. No crystallization was observed when the OIR is higher than 1:1. Suppression of (NH4)2SO4 crystal growth was also observed under high viscous sucrose/(NH4)2SO4 droplets at lower RH.


A Single-Molecule View of Genome Editing Proteins: Biophysical Mechanisms for TALEs and CRISPR/Cas9

Luke Cuculis and Charles M. Schroeder

Exciting new advances in genome engineering have unlocked the potential to radically alter the treatment of human disease. In this review, we discuss the application of single-molecule techniques to uncover the mechanisms behind two premier classes of genome editing proteins: transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated system (Cas). These technologies have facilitated a striking number of gene editing applications in a variety of organisms; however, we are only beginning to understand the molecular mechanisms governing the DNA editing properties of these systems. Here, we discuss the DNA search and recognition process for TALEs and Cas9 that have been revealed by recent single-molecule experiments.


Monday, September 11, 2017

Light Scattering By Optically-Trapped Vesicles Affords Unprecedented Temporal Resolution Of Lipid-Raft Dynamics

Liam Collard, David Perez-Guaita, Bayan H. A. Faraj, Bayden R. Wood, Russell Wallis, Peter W. Andrew & Andrew J. Hudson

A spectroscopic technique is presented that is able to identify rapid changes in the bending modulus and fluidity of vesicle lipid bilayers on the micrometer scale, and distinguish between the presence and absence of heterogeneities in lipid-packing order. Individual unilamellar vesicles have been isolated using laser tweezers and, by measuring the intensity modulation of elastic back-scattered light, changes in the biophysical properties of lipid bilayers were revealed. Our approach offers unprecedented temporal resolution and, uniquely, physical transformations of lipid bilayers can be monitored on a length scale of micrometers. As an example, the deformation of a membrane bilayer following the gel-to-fluid phase transition in a pure phospholipid vesicle was observed to take place across an interval of 54 ± 5 ms corresponding to an estimated full-width of only ~1 m°C. Dynamic heterogeneities in packing order were detected in mixed-lipid bilayers. Using a ternary mixture of lipids, the modulated-intensity profile of elastic back-scattered light from an optically-trapped vesicle revealed an abrupt change in the bending modulus of the bilayer which could be associated with the dissolution of ordered microdomains (i.e., lipid rafts). This occurred across an interval of 30 ± 5 ms (equivalent to ~1 m°C).


Optical trapping forces of a focused azimuthally polarized Bessel-Gaussian beam on a double-layered sphere

F. P. Wu, B. Zhang, Z. L. Liu, Y. Tang, N. Zhang

We calculate the trapping forces exerted by a highly focused Bessel-Gaussian beam on a double-layered sphere by means of vector diffraction integral, T-matrix method and Maxwell stress tensor integral. The Bessel-Gaussian beam is azimuthally polarized. Numerical results predicate that the double-layered sphere with air core can be stably trapped in three-dimensions. The trapping forces and efficiencies are dependent on the refraction index and size of the inner core. The trapping efficiency can be optimized by choosing the refraction indices of the inner core and outer layer. Our computational method can be easily modified for other laser beams and particles with arbitrary geometries and multilayers.


Charge Effects on the Efflorescence in Single Levitated Droplets

Gunter Hermann, Yan Zhang, Bernhard Wassermann, Henry Fischer, Marcel Quennet, and Eckart Rühl

The influence of electrical excess charges on the crystallization from supersaturated aqueous sodium chloride solutions is reported. This is accomplished by efflorescence studies on single levitated microdroplets using optical and electrodynamic levitation. Specifically, a strong increase in efflorescence humidity is observed as a function of the droplet’s negative excess charge, ranging up to −2.1 pC, with a distinct threshold behavior, increasing the relative efflorescence humidity, at which spontaneous nucleation occurs, from 44% for the neutral microparticle to 60%. These findings are interpreted by using molecular dynamics simulations for determining plausible structural patterns located near the particle surface that could serve as suitable precursors for the formation of critical clusters overcoming the nucleation barrier. These results, facilitating heterogeneous nucleation in the case of negatively charged microparticles, are compared to recent work on charge-induced nucleation of neat supercooled water, where a distinctly different nucleation behavior as a function of droplet charge has been observed.


Vinculin forms a directionally asymmetric catch bond with F-actin

Derek L. Huang, Nicolas A. Bax, Craig D. Buckley, William I. Weis, Alexander R. Dunn

Vinculin is an actin-binding protein thought to reinforce cell-cell and cell-matrix adhesions. However, how mechanical load affects the vinculin–F-actin bond is unclear. Using a single-molecule optical trap assay, we found that vinculin forms a force-dependent catch bond with F-actin through its tail domain, but with lifetimes that depend strongly on the direction of the applied force. Force toward the pointed (–) end of the actin filament resulted in a bond that was maximally stable at 8 piconewtons, with a mean lifetime (12 seconds) 10 times as long as the mean lifetime when force was applied toward the barbed (+) end. A computational model of lamellipodial actin dynamics suggests that the directionality of the vinculin–F-actin bond could establish long-range order in the actin cytoskeleton. The directional and force-stabilized binding of vinculin to F-actin may be a mechanism by which adhesion complexes maintain front-rear asymmetry in migrating cells.


Dynamics of microtubules: highlights of recent computational and experimental investigations

Valeri Barsegov, Jennifer Ross and Ruxandra Dima

Microtubules are found in most eukaryotic cells, with homologs in eubacteria and archea, and they have functional roles in mitosis, cell motility, intracellular transport, and the maintenance of cell shape. Numerous efforts have been expended over the last two decades to characterize the interactions between microtubules and the wide variety of microtubule associated proteins that control their dynamic behavior in cells resulting in microtubules being assembled and disassembled where and when they are required by the cell. We present the main findings regarding microtubule polymerization and depolymerization and review recent work about the molecular motors that modulate microtubule dynamics by inducing either microtubule depolymerization or severing. We also discuss the main experimental and computational approaches used to quantify the thermodynamics and mechanics of microtubule filaments.


Total momentum transfer produced by the photons of a multi-pass laser beam as an evident avenue for optical and mass metrology

Suren Vasilyan, Thomas Fröhlich, and Eberhard Manske
The use of the radiation pressure of a laser field, as an effect of the momentum transfer of the absorbed and re-emitted photons, suggests rather a complementary than an alternative possibility for metrology to generate calibration forces or to calibrate the optical power directly traceable to the International System of Units (SI). This paper reports a method and experimentally measured evidence on options to extend the effective use of radiation pressure for generating optical forces in the sub-microNewton (μN) range. Among other features and results presented, we emphasize the variability in controlling the accuracy of these forces through the proper utilization of the power of a multi-pass laser beam (semi- or completely) locked within confined systems. The direct measurements of these forces, augmented due to the partial or total momentum transfer of the photons of a multi-pass laser beam extended from several hundreds of picoNewton (pN) up to sub-μN range for the same power of laser source, are done by a precision force measurement system and compared with basic theoretical computations. We also discuss the opportunities to probe the fundamental physical limits associated with this method and to the considerable extent other competing contributing effects that might be regarded as sources of errors in metrological tasks.


Wednesday, September 6, 2017

Pausing kinetics dominates strand-displacement polymerization by reverse transcriptase

Omri Malik, Hadeel Khamis, Sergei Rudnizky, Ailie Marx, Ariel Kaplan

Reverse transcriptase (RT) catalyzes the conversion of the viral RNA into an integration-competent double-stranded DNA, with a variety of enzymatic activities that include the ability to displace a non-template strand concomitantly with polymerization. Here, using high-resolution optical tweezers to follow the activity of the murine leukemia Virus RT, we show that strand-displacement polymerization is frequently interrupted. Abundant pauses are modulated by the strength of the DNA duplex ∼8 bp ahead, indicating the existence of uncharacterized RT/DNA interactions, and correspond to backtracking of the enzyme, whose recovery is also modulated by the duplex strength. Dissociation and reinitiation events, which induce long periods of inactivity and are likely the rate-limiting step in the synthesis of the genome in vivo, are modulated by the template structure and the viral nucleocapsid protein. Our results emphasize the potential regulatory role of conserved structural motifs, and may provide useful information for the development of potent and specific inhibitors.


Piconewton-Scale Analysis of Ras-BRaf Signal Transduction with Single-Molecule Force Spectroscopy

Chae-Seok Lim, Cheng Wen, Yanghui Sheng, Guangfu Wang, Zhuan Zhou, Shiqiang Wang, Huaye Zhang, Anpei Ye, J. Julius Zhu

Intermolecular interactions dominate the behavior of signal transduction in various physiological and pathological cell processes, yet assessing these interactions remains a challenging task. Here, this study reports a single-molecule force spectroscopic method that enables functional delineation of two interaction sites (≈35 pN and ≈90 pN) between signaling effectors Ras and BRaf in the canonical mitogen-activated protein kinase (MAPK) pathway. This analysis reveals mutations on BRaf at Q257 and A246, two sites frequently linked to cardio-faciocutaneous syndrome, result in ≈10−30 pN alterations in Ras[BOND]BRaf intermolecular binding force. The magnitude of changes in Ras[BOND]BRaf binding force correlates with the size of alterations in protein affinity and in α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-sensitive glutamate receptor (-R)-mediated synaptic transmission in neurons expressing replacement BRaf mutants, and predicts the extent of learning impairments in animals expressing replacement BRaf mutants. These results establish single-molecule force spectroscopy as an effective platform for evaluating the piconewton-level interaction of signaling molecules and predicting the behavior outcome of signal transduction.


Higher-order micro-fiber modes for Escherichia coli manipulation using a tapered seven-core fiber

Qiangzhou Rong, Yi Zhou, Xunli Yin, Zhihua Shao, and Xueguang Qiao

Optical manipulation using optical micro- and nano-fibers has shown potential for controlling bacterial activities such as E. coli trapping, propelling, and binding. Most of these manipulations have been performed using the propagation of the fundamental mode through the fiber. However, along the maximum mode-intensity axis, the higher-order modes have longer evanescent field extensions and larger field amplitudes at the fiber waist than the fundamental mode, opening up new possibilities for manipulating E. coli bacteria. In this work, a compact seven-core fiber (SCF)-based micro-fiber/optical tweezers was demonstrated for trapping, propelling, and rotating E. coli bacteria using the excitation of higher-order modes. The diameter of the SCF taper was 4 µm at the taper waist, which was much larger than that of previous nano-fiber tweezers. The laser wavelength was tunable from 1500 nm to 1600 nm, simultaneously causing photophoretic force, gradient force, and scattering force. This work provides a new opportunity for better understanding optical manipulation using higher-order modes at the single-cell level.


Understanding and Reducing Photothermal Forces for the Fabrication of Au Nanoparticle Dimers by Optical Printing

Julian Gargiulo, Thomas Brick, Ianina L. Violi, Facundo C. Herrera, Toshihiko Shibanuma, Pablo Albella, Félix G. Requejo, Emiliano Cortés, Stefan A. Maier, and Fernando D. Stefani

Optical printing holds great potential to enable the use of the vast variety of colloidal nanoparticles (NPs) in nano- and microdevices and circuits. By means of optical forces, it enables the direct assembly of NPs, one by one, onto specific positions of solid surfaces with great flexibility of pattern design and no need of previous surface patterning. However, for unclear causes it was not possible to print identical NPs closer to each other than 300 nm. Here, we show that the repulsion restricting the optical printing of close by NPs arises from light absorption by the printed NPs and subsequent local heating. By optimizing heat dissipation, it is possible to reduce the minimum separation between NPs. Using a reduced graphene oxide layer on a sapphire substrate, we demonstrate for the first time the optical printing of Au—Au NP dimers. Modeling the experiments considering optical, thermophoretic, and thermo-osmotic forces we obtain a detailed understanding and a clear pathway for the optical printing fabrication of complex nano structures and circuits based on connected colloidal NPs.


A DNA-centered explanation of the DNA polymerase translocation mechanism

J. Ricardo Arias-Gonzalez

DNA polymerase couples chemical energy to translocation along a DNA template with a specific directionality while it replicates genetic information. According to single-molecule manipulation experiments, the polymerase-DNA complex can work against loads greater than 50 pN. It is not known, on the one hand, how chemical energy is transduced into mechanical motion, accounting for such large forces on sub-nanometer steps, and, on the other hand, how energy consumption in fidelity maintenance integrates in this non-equilibrium cycle. Here, we propose a translocation mechanism that points to the flexibility of the DNA, including its overstretching transition, as the principal responsible for the DNA polymerase ratcheting motion. By using thermodynamic analyses, we then find that an external load hardly affects the fidelity of the copying process and, consequently, that translocation and fidelity maintenance are loosely coupled processes. The proposed translocation mechanism is compatible with single-molecule experiments, structural data and stereochemical details of the DNA-protein complex that is formed during replication, and may be extended to RNA transcription.


Tuesday, September 5, 2017

Control of nucleus positioning in mouse oocytes

Maria Almonacid, Marie-EmilieTerret, Marie-Hélène Verlhac

The position of the nucleus in a cell can instruct morphogenesis in some cases, conveying spatial and temporal information and abnormal nuclear positioning can lead to disease. In oocytes from worm, sea urchin, frog and some fish, nucleus position regulates embryo development, it marks the animal pole and in Drosophila it defines the future dorso-ventral axis of the embryo and of the adult body plan. However, in mammals, the oocyte nucleus is centrally located and does not instruct any future embryo axis. Yet an off-center nucleus correlates with poor outcome for mouse and human oocyte development. This is surprising since oocytes further undergo two extremely asymmetric divisions in terms of the size of the daughter cells (enabling polar body extrusion), requiring an off-centering of their chromosomes. In this review we address not only the bio-physical mechanism controlling nucleus positioning via an actin-mediated pressure gradient, but we also speculate on potential biological relevance of nuclear positioning in mammalian oocytes and early embryos.


3D correlative single-cell imaging utilizing fluorescence and refractive index tomography

Mirjam Schürmann, Gheorghe Cojoc, Salvatore Girardo, Elke Ulbricht, Jochen Guck, Paul Müller

Cells alter the path of light, a fact that leads to well-known aberrations in single cell or tissue imaging. Optical diffraction tomography (ODT) measures the biophysical property that causes these aberrations, the refractive index (RI). ODT is complementary to fluorescence imaging and does not require any markers. The present study introduces RI and fluorescence tomography with optofluidic rotation (RAFTOR) of suspended cells, facilitating the segmentation of the 3D-correlated RI and fluorescence data for a quantitative interpretation of the nuclear RI. The technique is validated with cell phantoms and used to confirm a lower nuclear RI for HL60 cells. Furthermore, the nuclear inversion of adult mouse photoreceptor cells is observed in the RI distribution. The applications shown confirm predictions of previous studies and illustrate the potential of RAFTOR to improve our understanding of cells and tissues.


Moiré deflectometry-based position detection for optical tweezers

Ali Akbar Khorshad, S. Nader S. Reihani, and Mohammad Taghi Tavassoly

Optical tweezers have proven to be indispensable tools for pico-Newton range force spectroscopy. A quadrant photodiode (QPD) positioned at the back focal plane of an optical tweezers’ condenser is commonly used for locating the trapped object. In this Letter, for the first time, to the best of our knowledge, we introduce a moiré pattern-based detection method for optical tweezers. We show, both theoretically and experimentally, that this detection method could provide considerably better position sensitivity compared to the commonly used detection systems. For instance, position sensitivity for a trapped 2.17 μm polystyrene bead is shown to be 71% better than the commonly used QPD-based detection method. Our theoretical and experimental results are in good agreement.


Thermal tuning of spectral emission from optically trapped liquid-crystal droplet resonators

Alexandr Jonáš, Zdeněk Pilát, Jan Ježek, Silvie Bernatová, Tomáš Fořt, Pavel Zemánek, Mehdi Aas, and Alper Kiraz

Surfactant-stabilized emulsion droplets of liquid crystals (LCs) suspended in water and labeled with a fluorescent dye form active, anisotropic optofluidic microresonators. These microresonators can host whispering gallery modes (WGMs), high-quality morphology-dependent optical resonances that are supported due to the contrast of refractive index between the LC droplets and the surrounding aqueous medium. In addition, owing to the refractive index contrast, such LC emulsion droplets can be stably trapped in three dimensions using optical tweezers, enabling long-term investigation of their spectral characteristics. We explore various combinations of fluorescently dyed LC droplets and host liquid-surfactant systems and show that the WGM emission spectra of optical resonators based on optically trapped LC emulsion droplets can be largely and (almost) reversibly tuned by controlled changes of the ambient temperature. Depending on the actual range of temperature modulation and LC phase of the studied droplet, thermally induced effects can either lead to phase transitions in the LC droplets or cause modifications of their refractive index profile without changing their LC phase. Our results indicate feasibility of this approach for creating miniature thermally tunable sources of coherent light that can be manipulated and stabilized by optical forces.


Directional Optical Sorting of Silicon Nanoparticles

Daniil A. Shilkin, Evgeny V. Lyubin, Maxim R. Shcherbakov, Mikhail Lapine, and Andrey A. Fedyanin

Optical manipulation of nanoparticles is a topic of great practical importance, with applications in surface science, colloidal chemistry, microfluidics, biochemistry, and medicine. One of the major highlights of this topic is particle sorting, which serves to create monodisperse systems remotely and to separate particles of different composition and size. Here, we analyze optical forces acting on spherical silicon nanoparticles that exhibit high-quality Mie resonances and demonstrate the potential of optical sorting methods for these systems. In particular, we propose multidirectional static sorting of nanoparticles using noncollinear beams with different wavelengths, which results in all-optical separation into an angular spectrum of sizes. We also validate the proposed methods by considering the operation in the presence of Brownian motion and in the evanescent wave configuration.


Experiment study and FEM simulation on erythrocytes under linear stretching of optical micromanipulation

Ying Liu, Huadong Song, Panpan Zhu, Hao Lu, and Qi Tang

The elasticity of erythrocytes is an important criterion to evaluate the quality of blood. This paper presents a novel research on erythrocytes’ elasticity with the application of optical tweezers and the finite element method (FEM) during blood storage. In this work, the erythrocytes with different in vitro times were linearly stretched by trapping force using optical tweezers and the time dependent elasticity of erythrocytes was investigated. The experimental results indicate that the membrane shear moduli of erythrocytes increased with the increasing in vitro time, namely the elasticity was decreasing. Simultaneously, an erythrocyte shell model with two parameters (membrane thickness h and membrane shear modulus H) was built to simulate the linear stretching states of erythrocytes by the FEM, and the simulations conform to the results obtained in the experiment. The evolution process was found that the erythrocytes membrane thicknesses were decreasing. The analysis assumes that the partial proteins and lipid bilayer of erythrocyte membrane were decomposed during the in vitro preservation of blood, which results in thin thickness, weak bending resistance, and losing elasticity of erythrocyte membrane. This study implies that the FEM can be employed to investigate the inward mechanical property changes of erythrocyte in different environments, which also can be a guideline for studying the erythrocyte mechanical state suffered from different diseases.


Monday, September 4, 2017

Mode conversion enables optical pulling force in photonic crystal waveguides

Tongtong Zhu, Andrey Novitsky, Yongyin Cao, M. R. C. Mahdy, Lin Wang, Fangkui Sun, Zehui Jiang, and Weiqiang Ding

We propose a robust scheme to achieve optical pulling force using the guiding modes supported in a hollow core double-mode photonic crystal waveguide instead of the structured optical beams in free space investigated earlier. The waveguide under consideration supports both the 0th order mode with a larger forward momentum and the 1st order mode with a smaller forward momentum. When the 1st order mode is launched, the scattering by the object inside the waveguide results in the conversion from the 1st order mode to the 0th order mode, thus creating the optical pulling force according to the conservation of linear momentum. We present the quantitative agreement between the results derived from the mode conversion analysis and those from rigorous simulation using the finite-difference in the time-domain numerical method. Importantly, the optical pulling scheme presented here is robust and broadband with naturally occurred lateral equilibriums and has a long manipulation range. Flexibilities of the current configuration make it valuable for the optical force tailoring and optical manipulation operation, especially in microfluidic channel systems.

Investigation on the photophoretic lift force acting upon particles under light irradiation

Shuangling Dong, Yafei Liu

Considering the characteristics of photophoresis in the actual process, the viewpoint that particles in fluids will experience photophoretic lift force is proposed. The force is related to the particle motion with photophoresis but different from the traditional photophoretic force. Analysis shows that there are mainly two factors contributing to the lift force, one stems from the variation in the distribution of light intensity, the other originates from the rotational motion of particles in a direction which is perpendicular to the optical axis. The expression of the photophoretic lift force has been given in the study. The impacts of the force have been analyzed and validated by comparing with previous experimental results.


Single-molecule imaging and manipulation of biomolecular machines and systems

Ryota Iino, Tatsuya Iida, Akihiko Nakamura, Ei-ichiro Saita, Huijuan You, Yasushi Sako

Biological molecular machines support various activities and behaviors of cells, such as energy production, signal transduction, growth, differentiation, and migration.
We provide an overview of single-molecule imaging methods involving both small and large probes used to monitor the dynamic motions of molecular machines in vitro (purified proteins) and in living cells, and single-molecule manipulation methods used to measure the forces, mechanical properties and responses of biomolecules. We also introduce several examples of single-molecule analysis, focusing primarily on motor proteins and signal transduction systems.
Single-molecule analysis is a powerful approach to unveil the operational mechanisms both of individual molecular machines and of systems consisting of many molecular machines.


Exploring the Denatured State Ensemble by Single-Molecule Chemo-Mechanical Unfolding: The Effect of Force, Temperature, and Urea

Emily J.Guinn, SusanMarqusee

While it is widely appreciated that the denatured state of a protein is a heterogeneous conformational ensemble, there is still debate over how this ensemble changes with environmental conditions. Here, we use single-molecule chemo-mechanical unfolding, which combines force and urea using the optical tweezers, together with traditional protein unfolding studies to explore how perturbants commonly used to unfold proteins (urea, force, and temperature) affect the denatured-state ensemble. We compare the urea m-values, which report on the change in solvent accessible surface area for unfolding, to probe the denatured state as a function of force, temperature, and urea. We find that while the urea- and force-induced denatured states expose similar amounts of surface area, the denatured state at high temperature and low urea concentration is more compact. To disentangle these two effects, we use destabilizing mutations that shift the Tm and Cm. We find that the compaction of the denatured state is related to changing temperature as the different variants of acyl-coenzyme A binding protein have similar m-values when they are at the same temperature but different urea concentration. These results have important implications for protein folding and stability under different environmental conditions.


Optical Bessel beam illumination of a subwavelength prolate gold (Au) spheroid coated by a layer of plasmonic material: radiation force, spin and orbital torques

FG Mitri

The optical radiation force, spin and orbital torques exerted on a subwavelength prolate gold spheroid coated by a layer of plasmonic material with negative permittivity and illuminated by either a zeroth-order (non-vortex) or a first-order vector Bessel (vortex) beam are computed in the framework of the electric dipole approximation method. Calculations for the Cartesian components of the optical radiation force on a subwavelength spheroid with arbitrary orientation in space are performed, with emphasis on the order (or topological charge), half-cone angle of the beam, and the plasmonic layer thickness on- and off-resonance. A repulsive (pushing) force is predicted for the layered subwavelength prolate spheroid, on- and off-resonance along the direction of wave propagation. Moreover, the Cartesian components of the spin radiation torque are computed where a negative longitudinal spin torque component can arise, suggesting a rotational twist of the spheroid around its center of mass in either the counter-clockwise or the clockwise (negative) direction of spinning. In addition, the longitudinal component of the orbital radiation torque exhibits sign reversal, indicating a revolution around the beam axis in either the counter-clockwise or the clockwise directions. The results show that the plasmonic resonance strongly alters the force, spin and orbital torque components, causing major amplitude enhancements, signs twists, and complex distributions in the transverse plane.


Theoretical estimation of nonlinear optical force on dielectric spherical particles of arbitrary size under femtosecond pulsed excitation

Anita Devi and Arijit K. De

Experimental evidence indicates that high-repetition-rate ultrafast pulsed excitation is more efficient in optical trapping of dielectric nanoparticles as compared with continuous-wave excitation at the same average power. The physics behind the different nature of force under these two excitation conditions remained deceptive until quite recently when it was theoretically explained, in the dipole limit, as a combined effect of (1) repetitive instantaneous momentum transfer and (2) optical Kerr nonlinearity. The role of optical Kerr effect was theoretically studied for larger dielectric spherical particles, in the ray optics limit, also. However, a theoretical underpinning is yet to be established as to whether the effect of optical nonlinearity is omnipresent across different particle sizes, which we investigate here. Using localized approximation of generalized Lorenz-Mie theory, we theoretically analyze the nature of force (and potential) and provide a detailed comparative discussion between this generalized scattering formulation with dipole scattering formulation for dielectric nanoparticles.


Thursday, August 31, 2017

Substrate and Fano Resonance Effects on the Reversal of Optical Binding Force between Plasmonic Cube Dimers

M. R. C. Mahdy, Tianhang Zhang, Md. Danesh & Weiqiang Ding

The behavior of Fano resonance and the reversal of near field optical binding force of dimers over different substrates have not been studied so far. Notably, for particle clustering and aggregation, controlling the near filed binding force can be a key factor. In this work, we observe that if the closely located plasmonic cube homodimers over glass or high permittivity dielectric substrate are illuminated with plane wave, no reversal of lateral optical binding force occurs. But if we apply the same set-up over a plasmonic substrate, stable Fano resonance occurs along with the reversal of near field lateral binding force. It is observed that during such Fano resonance, stronger coupling occurs between the dimers and plasmonic substrate along with the strong enhancement of the substrate current. Such binding force reversals of plasmonic cube dimers have been explained based on the observed unusual behavior of optical Lorentz force during the induced stronger Fano resonance and the dipole-dipole resonance. Although previously reported reversals of near field optical binding forces were highly sensitive to particle size/shape (i.e. for heterodimers) and inter-particle distance, our configuration provides much relaxation of those parameters and hence could be verified experimentally with simpler experimental set-ups.


Saturated PID Control for the Optical Manipulation of Biological Cells

Mingyang Xie ; Xiaojian Li ; Yong Wang ; Yunhui Liu ; Dong Sun

This brief presents a saturated PID control scheme for cell manipulation by using robot-aided optical tweezers, with consideration of both translational and rotational control of the cell. By incorporating saturation functions into a PID controller and utilizing a velocity observer, cell position and orientation can asymptotically converge to the desired values. The proposed control approach also guarantees that the trapped cell can always stay within a small neighborhood around the centroid of the optical trap, thereby preventing the cell from escaping by optical trapping. The controller does not require the use of accurate dynamic model parameters, and hence can be implemented easily. Utilizing a velocity observer, velocity measurement by directly differentiating the measured position of the cell is not needed, which benefits noise reduction and system stability. The asymptotic stability of the closed-loop system is analyzed using Lyapunov's direct method. Experimental results are presented to demonstrate the effectiveness of the proposed approach.


Ultrahigh Purcell Factor, Improved Sensitivity, and Enhanced Optical Force in Dielectric Bowtie Whispering-Gallery-Mode Resonators

Qijing Lu ; Xiaogang Chen ; Hongqin Yang ; Xiang Wu ; Shusen Xie

We propose and theoretically investigate an all-dielectric bowtie whispering-gallery-mode (WGM) resonator which consists of two tip-to-tip coupled semiconductor nanorings with triangular cross section separated by low refractive index material gap. Mode splitting of symmetric WGM and antisymmetric WGM is observed and analyzed. Due to extremely field enhancements by the “slot” and “tip” effects, strong localization of light in the gap of symmetric WGMs leads to ultrasmall modal volume of 0.3μm3 . This value is two orders of magnitude smaller than that in toroidal microresonator which is the typical WGM microcavity with tight mode confinement. Importantly, large amount of light confined in the gap in bowtie WGM resonator not only suppresses the radiation loss (radiation-loss-related quality factor is above 108), but also makes it as an ideal platform for Purcell effect enhancement, ultrasensitive sensing, and optical trapping of nanoparticles. Calculation results show that Purcell factor can reach to 108 at room temperature when assuming quantum dots or atoms placed in the gap. Refractive index sensitivity is improved to 700 nm/RIU as compared with conventional slot waveguide with rectangular cross section. The optical gradient force is greatly enhanced and allows efficient trapping of single nanoscale particle with diameter of 5 nm even at a relatively large gap of 100 nm.


Heterogeneous interface adsorption of colloidal particles

Dong Woo Kang, Jin Hyun Lim and Bum Jun Park
The heterogeneous adsorption behaviors of charged colloidal particles to oil–water interfaces were quantitatively and statistically investigated. Using optical laser tweezers, the particles in a sessile water drop formed in an oil phase were laterally translated toward the slope of the oil–water interface and their attachment to the interface was attempted. The adsorption probability was found to logarithmically decrease as the ionic strength decreased and to depend on the holding time during which an optically trapped particle was held at the position closest to the interface. Non-unity of the adsorption probability at particular salt concentrations and the holding time dependence offer an important clue that the particle adsorption to the interface is not deterministic but stochastic. The stochastic adsorption process can be attributed to the surface heterogeneity of colloidal particles that consequently leads to changes in the electrostatic interactions between the particles and the interface. We also demonstrated that the salt dependence on the adsorption properties of the particles, as measured by optical laser tweezers, was consistent with their bulk behaviors with regard to the stability of particle-stabilized emulsions. Furthermore, we revealed the gravity-induced spontaneous adsorption of the particles to the interface under conditions of sufficiently strong ionic strength.


Detection and characterization of chemical aerosol using laser-trapping single-particle Raman spectroscopy

Aimable Kalume, Leonid A. Beresnev, Joshua Santarpia, and Yong-Le Pan

Detection and characterization of the presence of chemical agent aerosols in various complex atmospheric environments is an essential defense mission. Raman spectroscopy has the ability to identify chemical molecules, but there are limited numbers of photons detectable from single airborne aerosol particles as they are flowing through a detection system. In this paper, we report on a single-particle Raman spectrometer system that can measure strong spontaneous, stimulated, and resonance Raman spectral peaks from a single laser-trapped chemical aerosol particle, such as a droplet of the VX nerve agent chemical simulant diethyl phthalate. Using this system, time-resolved Raman spectra and elastic scattered intensities were recorded to monitor the chemical properties and size variation of the trapped particle. Such a system supplies a new approach for the detection and characterization of single airborne chemical aerosol particles.


Chirality and Chiroptical Effects in Metal Nanostructures: Fundamentals and Current Trends

Joel T. Collins, Christian Kuppe, David C. Hooper, Concita Sibilia, Marco Centini, Ventsislav K. Valev

Throughout the 19th and 20th century, chirality has mostly been associated with chemistry. However, while chirality can be very useful for understanding molecules, molecules are not well suited for understanding chirality. Indeed, the size of atoms, the length of molecular bonds and the orientations of orbitals cannot be varied at will. It is therefore difficult to study the emergence and evolution of chirality in molecules, as a function of geometrical parameters. By contrast, chiral metal nanostructures offer an unprecedented flexibility of design. Modern nanofabrication allows chiral metal nanoparticles to tune the geometric and optical chirality parameters, which are key for properties such as negative refractive index and superchiral light. Chiral meta/nano-materials are promising for numerous technological applications, such as chiral molecular sensing, separation and synthesis, super-resolution imaging, nanorobotics, and ultra-thin broadband optical components for chiral light. This review covers some of the fundamentals and highlights recent trends. We begin by discussing linear chiroptical effects. We then survey the design of modern chiral materials. Next, the emergence and use of chirality parameters are summarized. In the following part, we cover the properties of nonlinear chiroptical materials. Finally, in the conclusion section, we point out current limitations and future directions of development.


Wednesday, August 30, 2017

PFMCal : Photonic force microscopy calibration extended for its application in high-frequency microrheology

A. Butykai, P. Domínguez-García, F. M.Mor, R. Gaál, L. Forró, S. Jeney
The present document is an update of the previously published MatLab code for the calibration of optical tweezers in the high-resolution detection of the Brownian motion of non-spherical probes [1]. In this instance, an alternative version of the original code, based on the same physical theory [2], but focused on the automation of the calibration of measurements using spherical probes, is outlined. The new added code is useful for high-frequency microrheology studies, where the probe radius is known but the viscosity of the surrounding fluid maybe not. This extended calibration methodology is automatic, without the need of a user’s interface. A code for calibration by means of thermal noise analysis [3] is also included; this is a method that can be applied when using viscoelastic fluids if the trap stiffness is previously estimated [4]. The new code can be executed in MatLab and using GNU Octave.


Manipulating Living Cells to Construct a 3D Single-Cell Assembly without an Artificial Scaffold

Aoi Yoshida, Shoto Tsuji, Hiroaki Taniguchi, Takahiro Kenmotsu, Koichiro Sadakane and Kenichi Yoshikawa

Artificial scaffolds such as synthetic gels or chemically-modified glass surfaces that have often been used to achieve cell adhesion are xenobiotic and may harm cells. To enhance the value of cell studies in the fields of regenerative medicine and tissue engineering, it is becoming increasingly important to create a cell-friendly technique to promote cell–cell contact. In the present study, we developed a novel method for constructing stable cellular assemblies by using optical tweezers in a solution of a natural hydrophilic polymer, dextran. In this method, a target cell is transferred to another target cell to make cell–cell contact by optical tweezers in a culture medium containing dextran. When originally non-cohesive cells are held in contact with each other for a few minutes under laser trapping, stable cell–cell adhesion is accomplished. This method for creating cellular assemblies in the presence of a natural hydrophilic polymer may serve as a novel next-generation 3D single-cell assembly system with future applications in the growing field of regenerative medicine.


Multiple particles 3D trap based on all-fiber Bessel optical probe

Yaxun Zhang, Xiaoyun Tang, Yu Zhang, Zhihai Liu, Enming Zhao, Xinghua Yang, Jianzhong Zhang, Jun Yang, Libo Yuan

We propose and demonstrate an all-fiber Bessel optical tweezers for multiple microparticles (yeast cells) 3 dimensional trap. To the best knowledge of us, it is the first time to achieve the 3 dimensional stable non-contact multiple microparticles optical traps with long distance intervals by using a single all-fiber probe. The Bessel beam is produced by splicing coaxially a single mode fiber and a step index multimode fiber. The convergence of the output Bessel beam is performed by molding the tip of the multimode fiber into a special semi-ellipsoid shape. The effective trapping range of the all-fiber probe is 0 to 60μm, which is much longer than normal single fiber optical tweezers probes. The all-fiber Bessel optical probe is convenient to integrate and suitable for the lab on the chip. The structure of this fiber probe is simple, high-precision, low-cost, and small size, which provides new development for biological cells experiment and operation.


Ultra-high Q/V hybrid cavity for strong light-matter interaction

Donato Conteduca, Christopher Reardon, Mark G. Scullion, Francesco Dell’Olio, Mario N. Armenise, Thomas F. Krauss, and Caterina Ciminelli

The ability to confine light at the nanoscale continues to excite the research community, with the ratio between quality factor Q and volume V, i.e., the Q/V ratio, being the key figure of merit. In order to achieve strong light-matter interaction, however, it is important to confine a lot of energy in the resonant cavity mode. Here, we demonstrate a novel cavity design that combines a photonic crystal nanobeam cavity with a plasmonic bowtie antenna. The nanobeam cavity is optimised for a good match with the antenna and provides a Q of 1700 and a transmission of 90%. Combined with the bowtie, the hybrid photonic-plasmonic cavity achieves a Q of 800 and a transmission of 20%, both of which remarkable achievements for a hybrid cavity. The ultra-high Q/V of the hybrid cavity is of order of 106 (λ/n)−3, which is comparable to the state-of-the-art of photonic resonant cavities. Based on the high Q/V and the high transmission, we demonstrate the strong efficiency of the hybrid cavity as a nanotweezer for optical trapping. We show that a stable trapping condition can be achieved for a single 200 nm Au bead for a duration of several minutes (ttrap > 5 min) and with very low optical power (Pin = 190 μW).


Non-conservative optical forces

Sergey Sukhov and Aristide Dogariu

Undoubtedly, laser tweezers are the most recognized application of optically induced mechanical action. Their operation is usually described in terms of conservative forces originating from intensity gradients. However, the fundamental optical action on matter is non-conservative. We will review different manifestations of non-conservative optical forces (NCF) and discuss their dependence on the specific spatial properties of optical fields that generate them. New developments relevant to the NCF such as tractor beams and transversal forces are also discussed.


Tuesday, August 29, 2017

Displacement and localisation of a transparent nanosphere by light-pressure forces in the field of a focused laser beam

A.A. Afanas'ev and D.V. Novitsky

We report the results of a numerical simulation of the Langevin equation describing the motion of a transparent nanosphere under the action of a resulting light-pressure force in the field of a continuous focused Gaussian laser beam. The conditions for the localisation of the nanosphere near the focal waist of the beam focused by the lens are determined. An analytical solution of the approximate (truncated) equation of motion is found, which almost exactly coincides with the results of the numerical simulation of the initial equation.


Direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips

Jonathan W Driver, Elisabeth A Geyer, Megan E Bailey, Luke M Rice, Charles L Asbury

Disassembling microtubules can generate movement independently of motor enzymes, especially at kinetochores where they drive chromosome motility. A popular explanation is the ‘conformational wave’ model, in which protofilaments pull on the kinetochore as they curl outward from a disassembling tip. But whether protofilaments can work efficiently via this spring-like mechanism has been unclear. By modifying a previous assay to use recombinant tubulin and feedback-controlled laser trapping, we directly demonstrate the spring-like elasticity of curling protofilaments. Measuring their mechanical work output suggests they carry ~25% of the energy of GTP hydrolysis as bending strain, enabling them to drive movement with efficiency similar to conventional motors. Surprisingly, a β-tubulin mutant that dramatically slows disassembly has no effect on work output, indicating an uncoupling of disassembly speed from protofilament strain. These results show the wave mechanism can make a major contribution to kinetochore motility and establish a direct approach for measuring tubulin mechano-chemistry.


Multibeam interferometric optical tweezers

Mohammadbagher Mohammadnezhad; Abdollah Hassanzadeh

Using the interference of N collimated laser beams, optical lattices with N -fold rotational symmetry are generated over the interface of two semi-infinite dielectric media. The interaction of small dielectric particles with these interference patterns is investigated using Rayleigh approximation. The polarization state of the interfering beams considerably influences the interference patterns and potential landscapes. Therefore, both parallel and perpendicular polarized interfering beams are considered and the corresponding potential profiles are compared and analyzed. We also study how the number of interfering waves, incident and azimuth angles, and initial phases of the incident beams influence optical lattices and potential profiles. It is found that the ring-shaped patterns with good confinement properties can be achieved by increasing the number of incident beams. In addition, by increasing the number of incident beams one can make an optical trap with sharper intensity gradient and deeper potential well, which is an advantage for trapping small Rayleigh particles. The lattices resulting from the interference of N incident waves with different incident angles are also investigated. Furthermore, the effects of changing the azimuth angles between two adjacent incident wave vectors on the intensity patterns are studied. The proposed configuration and the numerical results can find numerous applications in particle arrangement, particle sorting, and the creation of quasicrystals. We believe that interference approaches have many potential capabilities for molding light wavefronts and creating multiple traps with sophisticated patterns.


In Vivo Manipulation of Single Biological Cells With an Optical Tweezers-Based Manipulator and a Disturbance Compensation Controller

Xiaojian Li ; Chichi Liu ; Shuxun Chen ; Yong Wang ; Shuk Han Cheng ; Dong Sun

In vivo manipulation of biological cells has attracted considerable attention in recent years. This process is particularly useful for precision medicine, such as cancer target therapy. Robotics technology is becoming necessary to stably and effectively manipulate and control single target cells in a complex in vivo environment. This paper presents a robot-aided optical tweezers-based manipulation technology that serves a function in the transport of single biological cells in vivo. An enhanced disturbance compensation controller is developed to minimize the effect of fluids (e.g., blood flow) on the cell. The method has exhibited advantages of flexibility in adjusting cell tracking trajectory online and the capability to minimize steady-state error and eliminate overshoot. Simulations and experiments of tracking single target cells in living zebrafish embryos have demonstrated the effectiveness of the proposed approach in a dynamic in vivo environment.


An early mechanical coupling of planktonic bacteria in dilute suspensions

Simon Sretenovic, Biljana Stojković, Iztok Dogsa, Rok Kostanjšek, Igor Poberaj & David Stopar

It is generally accepted that planktonic bacteria in dilute suspensions are not mechanically coupled and do not show correlated motion. The mechanical coupling of cells is a trait that develops upon transition into a biofilm, a microbial community of self-aggregated bacterial cells. Here we employ optical tweezers to show that bacteria in dilute suspensions are mechanically coupled and show long-range correlated motion. The strength of the coupling increases with the growth of liquid bacterial culture. The matrix responsible for the mechanical coupling is composed of cell debris and extracellular polymer material. The fragile network connecting cells behaves as viscoelastic liquid of entangled extracellular polymers. Our findings point to physical connections between bacteria in dilute bacterial suspensions that may provide a mechanistic framework for understanding of biofilm formation, osmotic flow of nutrients, diffusion of signal molecules in quorum sensing, or different efficacy of antibiotic treatments at low and high bacterial densities.


Monday, August 28, 2017

Drying-mediated optical assembly of silica spheres in a symmetrical metallic waveguide structure

Tian Xu, Cheng Yin, XueFen Kan, TingChao He, Qingbang Han, Zhuangqi Cao, and Xianfeng Chen

We describe the optical trapping application of a simple metallic slab optical waveguide structure, and demonstrate the influence of the excited guided modes on the aggregation behavior of silica particles during the irreversible evaporation process. Periodic horizontal stripes are formed by the highly ordered assemblies of the silica spheres, which is explained via the interference effect between the forward propagating modes and its reflection at the solvent surface. Particularly, several layers consisting of high-density particles are discernible in the stripe zones due to the optical binding, while no particles locate between these stripes. Completely different from the self-assembly patterns in the evaporating solvent without excitation of optical modes, this Letter demonstrates the versatility in the possible patterns of the optical assembly by a coupling waveguide with more complex structures.


Biphasic Effect of Profilin Impacts the Formin mDia1 Force-Sensing Mechanism in Actin Polymerization

Hiroaki Kubota, Makito Miyazaki, Taisaku Ogawa, Togo Shimozawa, Kazuhiko Kinosita Jr., Shin’ichi Ishiwata

Formins are force-sensing proteins that regulate actin polymerization dynamics. Here, we applied stretching tension to individual actin filaments under the regulation of formin mDia1 to investigate the mechanical responses in actin polymerization dynamics. We found that the elongation of an actin filament was accelerated to a greater degree by stretching tension for ADP-G-actin than that for ATP-G-actin. An apparent decrease in the critical concentration of G-actin was observed, especially in ADP-G-actin. These results on two types of G-actin were reproduced by a simple kinetic model, assuming the rapid equilibrium between pre- and posttranslocated states of the formin homology domain two dimer. In addition, profilin concentration dramatically altered the force-dependent acceleration of actin filament elongation, which ranged from twofold to an all-or-none response. Even under conditions in which actin depolymerization occurred, applications of a several-piconewton stretching tension triggered rapid actin filament elongation. This extremely high force-sensing mechanism of mDia1 and profilin could be explained by the force-dependent coordination of the biphasic effect of profilin; i.e., an acceleration effect masked by a depolymerization effect became dominant under stretching tension, negating the latter to rapidly enhance the elongation rate. Our findings demonstrate that the biphasic effect of profilin is controlled by mechanical force, thus expanding the function of mDia1 as a mechanosensitive regulator of actin polymerization.


Quantification of Chemical and Mechanical Effects on the Formation of the G-Quadruplex and i-Motif in Duplex DNA

Sangeetha Selvam, Shankar Mandal, and Hanbin Mao

The formation of biologically significant tetraplex DNA species, such as G-quadruplexes and i-motifs, is affected by chemical (ions and pH) and mechanical [superhelicity (σ) and molecular crowding] factors. Because of the extremely challenging experimental conditions, the relative importance of these factors on tetraplex folding is unknown. In this work, we quantitatively evaluated the chemical and mechanical effects on the population dynamics of DNA tetraplexes in the insulin-linked polymorphic region using magneto-optical tweezers. By mechanically unfolding individual tetraplexes, we found that ions and pH have the largest effects on the formation of the G-quadruplex and i-motif, respectively. Interestingly, superhelicity has the second largest effect followed by molecular crowding conditions. While chemical effects are specific to tetraplex species, mechanical factors have generic influences. The predominant effect of chemical factors can be attributed to the fact that they directly change the stability of a specific tetraplex, whereas the mechanical factors, superhelicity in particular, reduce the stability of the competing species by changing the kinetics of the melting and annealing of the duplex DNA template in a nonspecific manner. The substantial dependence of tetraplexes on superhelicity provides strong support that DNA tetraplexes can serve as topological sensors to modulate fundamental cellular processes such as transcription.


Recent advances in studying single bacteria and biofilm mechanics

Even C, Marlière C, Ghigo JM, Allain JM, Marcellan A, Raspaud E

Bacterial biofilms correspond to surface-associated bacterial communities embedded in hydrogel-like matrix, in which high cell density, reduced diffusion and physico-chemical heterogeneity play a protective role and induce novel behaviors. In this review, we present recent advances on the understanding of how bacterial mechanical properties, from single cell to high-cell density community, determine biofilm tri-dimensional growth and eventual dispersion and we attempt to draw a parallel between these properties and the mechanical properties of other well-studied hydrogels and living systems.


Effect of directional pulling on mechanical protein degradation by ATP-dependent proteolytic machines

Adrian O. Olivares, Hema Chandra Kotamarthi, Benjamin J. Stein, Robert T. Sauer and Tania A. Baker

AAA+ proteases and remodeling machines couple hydrolysis of ATP to mechanical unfolding and translocation of proteins following recognition of sequence tags called degrons. Here, we use single-molecule optical trapping to determine the mechanochemistry of two AAA+ proteases, Escherichia coli ClpXP and ClpAP, as they unfold and translocate substrates containing multiple copies of the titinI27 domain during degradation initiated from the N terminus. Previous studies characterized degradation of related substrates with C-terminal degrons. We find that ClpXP and ClpAP unfold the wild-type titinI27 domain and a destabilized variant far more rapidly when pulling from the N terminus, whereas translocation speed is reduced only modestly in the N-to-C direction. These measurements establish the role of directionality in mechanical protein degradation, show that degron placement can change whether unfolding or translocation is rate limiting, and establish that one or a few power strokes are sufficient to unfold some protein domains.


Light-driven micro- and nanomotors for environmental remediation

M. Safdar, J. Simmchen and J. Jänis

Synthetic micro- and nanomotors (MNMs) have emerged as a vibrant research field in multidisciplinary nanotechnology with proof-of-concept applications in various disciplines. In this tutorial review, an overview of the latest achievements towards light-driven MNMs is given and their propulsion mechanisms are introduced. The focus of the paper is on the autonomously propelled MNMs that exploit light-induced physical effects or chemical reactions. Light-induced body deformation, as a completely different, nature inspired concept that is found mostly in soft, polymeric MNMs, is also reviewed. In the end, a few applications of photocatalytic and light-driven MNMs for environmental remediation are presented and their potential is critically discussed.


Friday, August 25, 2017

Controlled shaping of lipid vesicles in a microfluidic diffusion chamber

M. Mally, B. Božič, S. Vrhovec Hartman, U. Klančnik, M. Mur, S. Svetina and J. Derganc

Synthetic lipid vesicles represent an important model system for studying membrane processes, which often depend on membrane shape, but controlled shaping of vesicles remains a challenging experimental task. Here, we present a novel method for shaping giant lipid vesicles by independently regulating osmotic conditions and the concentration of membrane-shaping molecules, which intercalate into the membrane and drive membrane bending. The method is based on the microfluidic diffusion chamber, where the solution around the vesicles can be repeatedly exchanged solely by diffusion, without any hydrodynamic flow that could deform the membrane. By using lipopolysaccharide (LPS) as a vesicle shape-modifying molecule, we demonstrate controlled and reversible transformations across three shape classes, from invaginated to evaginated vesicles. We show that extensive shape transformations can lead to shapes that are assumed to comprise narrow membrane necks that hinder equilibration of the membrane and the vesicle interior. All the observed shapes are in good agreement with the predictions of the area-difference-elasticity model applied to the vesicles that were denser than their surrounding solution. Our results validate the microfluidic diffusion chamber as a universal framework for membrane shaping that could also pave the way towards controlled fabrication of synthetic membranes resembling cell-compartments with large surface-to-volume ratios.


Using back focal plane interferometry to probe the influence of Zernike aberrations in optical tweezers

Thomas F. Dixon, Lachlan W. Russell, Ana Andres-Arroyo, and Peter J. Reece

We experimentally investigate the influence of geometric aberrations in optical tweezers using back focal plane interferometry. We found that the introduction of coma aberrations causes significant modification to the Brownian motion of the trapped particle, producing an apparent cross-coupling between the in-plane aberrated axis and the weaker propagation axis. This coupling is evidenced by the emergence of a second dominant low frequency Lorentzian feature in the position power spectral density. The effect on Brownian motion was confirmed using a secondary unaberrated probe beam, ruling out the possibility of systematic optical effects related to the detection system.


Automated Transportation of Biological Cells for Multiple Processing Steps in Cell Surgery

Hao Yang ; Xiangpeng Li ; Yunhui Liu ; Dong Sun

Most studies on automated cell transportation are single-task oriented. Results from these investigations hardly meet the increasing demand for emerging cell surgery operations that usually require a series of manipulation tasks with multiple processing steps. In this paper, automated cell transportation to accomplish a multistep process in cell surgery was investigated. A novel control system that can manipulate grouped cells to move into different task regions sequentially and continuously without interruption was developed based on a robot-aided optical tweezers manipulation system. A potential field-based controller was designed to achieve multistep processing control, where the new concepts of contractive coalition and switching region were incorporated into tweezers-cell coalition. The success of this controller lies in simultaneously controlling the positions of the optical tweezers, trapping multiple cells effectively, and avoiding collisions in a unified manner. Simulations and experiments of transferring a group of cells to a number of task regions were performed to demonstrate the effectiveness of the proposed approach.


Optomechanical soft metamaterials

Xiangjun Peng, Wei He, Yifan Liu, Fengxian Xin, Tian Jian Lu

We present a new type of optomechanical soft metamaterials, which is different from conventional mechanical metamaterials, in that they are simple isotropic and homogenous materials without resorting to any complex nano/microstructures. This metamaterial is unique in the sense that its responses to uniaxial forcing can be tailored by programmed laser inputs to manifest different nonlinear constitutive behaviors, such as monotonic, S-shape, plateau, and non-monotonic snapping performance. To demonstrate the novel metamaterial, a thin sheet of soft material impinged by two counterpropagating lasers along its thickness direction and stretched by an in-plane tensile mechanical force is considered. A theoretical model is formulated to characterize the resulting optomechanical behavior of the thin sheet by combining the nonlinear elasticity theory of soft materials and the optical radiation stress theory. The optical radiation stresses predicted by the proposed model are validated by simulations based on the method of finite elements. Programmed optomechanical behaviors are subsequently explored using the validated model under different initial sheet thicknesses and different optical inputs, and the first- and second-order tangential stiffness of the metamaterial are used to plot the phase diagram of its nonlinear constitutive behaviors. The proposed optomechanical soft metamaterial shows great potential in biological medicine, microfluidic manipulation, and other fields.


Stochastic analysis of time series for the spatial positions of particles trapped in optical tweezers

S. M. Mousavi, S. N. Seyed Reihani, G. Anvari, M. Anvari, H. G. Alinezhad & M. Reza Rahimi Tabar

We propose a nonlinear method for the analysis of the time series for the spatial position of a bead trapped in optical tweezers, which enables us to reconstruct its dynamical equation of motion. The main advantage of the method is that all the functions and parameters of the dynamics are determined directly (non-parametrically) from the measured series. It also allows us to determine, for the first time to our knowledge, the spatial-dependence of the diffusion coefficient of a bead in an optical trap, and to demonstrate that it is not in general constant. This is in contrast with the main assumption of the popularly-used power spectrum calibration method. The proposed method is validated via synthetic time series for the bead position with spatially-varying diffusion coefficients. Our detailed analysis of the measured time series reveals that the power spectrum analysis overestimates considerably the force constant.


Dynamic fatigue measurement of human erythrocytes using dielectrophoresis

Qiang Y, Liu J, Du E

Erythrocytes must undergo severe deformation to pass through narrow capillaries and submicronic splenic slits for several hundred thousand times in their normal lifespan. Studies of erythrocyte biomechanics have been mainly focused on cell deformability and rheology measured from a single application of stress and mostly under a static or quasi-static state using classical biomechanical techniques, such as optical tweezers and micropipette aspiration. Dynamic behavior of erythrocytes in response to cyclic stresses that contributes to the membrane failure in blood circulation is not fully understood. This paper presents a new experimental method for dynamic fatigue analysis of erythrocytes, using amplitude modulated electrokinetic force field in a microfluidic platform. We demonstrate the capability of this new technique using a low cycle fatigue analysis of normal human erythrocytes and ATP-depleted erythrocytes. Cyclic tensile stresses are generated to induce repeated uniaxial stretching and extensional recovery of single erythrocytes. Results of morphological and biomechanical parameters of individually tracked erythrocytes show strong correlations with the number of the loading cycles. Under a same strength of electric field, after 180 stress cycles, for normal erythrocytes, maximum stretch ratio decreases from 3.80 to 2.86, characteristic time of cellular extensional recovery increases from 0.16s to 0.37s, membrane shear viscosity increases from 1.0(µN/m)s to 1.6(µN/m)s. Membrane deformation in a small number of erythrocytes becomes irreversible after large deformation for about 200 cyclic loads. ATP-depleted cells show similar trends in decreased deformation and increased characteristic time with the loading cycles. These results show proof of concept of the new microfluidics technique for dynamic fatigue analysis of human erythrocytes.


Thursday, August 24, 2017

One-dimensional photonic crystals bound by light

Liyong Cui, Xiao Li, Jun Chen, Yongyin Cao, Guiqiang Du, and Jack Ng
Through rigorous simulations, the light scattering induced optical binding of one-dimensional (1D) dielectric photonic crystals is studied. The optical forces corresponding to the pass band, band gap, and band edge are qualitatively different. It is shown that light can induce self-organization of dielectric slabs into stable photonic crystals, with its lower band edge coinciding with the incident light frequency. Incident light at normal and oblique incidence and photonic crystals with parity-time symmetry are also considered.


Trapping two types of particles using a focused partially coherent circular edge dislocations beam

Hanghang Zhang, Jinhong Li, Ke Cheng, Meiling Duan, Zhifang Feng

A focused partially coherent circular edge dislocations beam used to trap Rayleigh dielectric sphere with different refractive indices is studied. The dependence of radiation forces on the number of circular edge dislocations p, the spatial correlation length σ0, relative refractive index nr, and particle radius a are analyzed and illustrated by numerical examples. It is shown that the focused partially coherent circular edge dislocations beam can be used to trap high index of refraction particles at focus F and bright ring R2, and simultaneously to capture low index of refraction particles at dark ring R1. It is much easier to capture the high index of refraction particles at focus F and the low index of refraction particles at dark ring as for the larger number of circular edge dislocations p and the spatial correlation length σ0, therefore it is necessary to optimally choose on p and σ0 for obtaining an optimal optical guiding. The ranges of the radius for two types of particles stably captured also have been determined. The obtained results are useful for analyzing the trapping efficiency of circular edge dislocations beams applied in micromanipulation technology.


Chiral Rayleigh particles discrimination in dynamic dual optical traps

Luis Carretero, Pablo Acebal, Salvador Blaya

A chiral optical conveyor belt for enantiomeric separation of nanoparticles is numerically demonstrated by using different types of counter propagating elliptical Laguerre Gaussian beams with different beam waist and topological charge. The analysis of chiral resolution has been made for particles immersed in water demonstrating that in the analyzed conditions one type of enantiomer is trapped in a deep potential and the others are transported by the chiral conveyor toward another trap located in a different geometrical region.


Light Shaping with Holography, GPC and Holo-GPC

Andrew Bañas, Jesper Glückstad

Light shaping techniques based on phase-only modulation offer multiple advantages over amplitude modulation. This review examines and compares the merits of two phase modulation techniques; phase-only computer generated holography and Generalized Phase Contrast (GPC). Both techniques are briefly presented while recent developments in GPC will also be covered. Furthermore, novel hybrid schemes that inherit merits from both holography and GPC are also covered. In particular, our most recent technique coined “Holo-GPC” will be discussed in addition to earlier hybrid techniques. We will discuss how Holo-GPC utilizes the simplicity of GPC in forming well-defined speckle-free shapes and the versatility of holography in distributing these shaped beams over an extended 3D volume. To conclude, we cite applications where the combined strengths of the two photon-efficient phase-only light shaping techniques open new possibilities.


A stretched conformation of DNA with a biological role?

Niklas Bosaeus, Anna Reymer, Tamás Beke-Somfai, Tom Brown, Masayuki Takahashi, Pernilla Wittung-Stafshede, Sandra Rocha and Bengt Nordén

We have discovered a well-defined extended conformation of double-stranded DNA, which we call Σ-DNA, using laser-tweezers force-spectroscopy experiments. At a transition force corresponding to free energy change ΔG = 1·57 ± 0·12 kcal (mol base pair)−1 60 or 122 base-pair long synthetic GC-rich sequences, when pulled by the 3′−3′ strands, undergo a sharp transition to the 1·52 ± 0·04 times longer Σ-DNA. Intriguingly, the same degree of extension is also found in DNA complexes with recombinase proteins, such as bacterial RecA and eukaryotic Rad51. Despite vital importance to all biological organisms for survival, genome maintenance and evolution, the recombination reaction is not yet understood at atomic level. We here propose that the structural distortion represented by Σ-DNA, which is thus physically inherent to the nucleic acid, is related to how recombination proteins mediate recognition of sequence homology and execute strand exchange. Our hypothesis is that a homogeneously stretched DNA undergoes a ‘disproportionation’ into an inhomogeneous Σ-form consisting of triplets of locally B-like perpendicularly stacked bases. This structure may ensure improved fidelity of base-pair recognition and promote rejection in case of mismatch during homologous recombination reaction. Because a triplet is the length of a gene codon, we speculate that the structural physics of nucleic acids may have biased the evolution of recombinase proteins to exploit triplet base stacks and also the genetic code.


Wednesday, August 23, 2017

Levitation and propulsion of a Mie-resonance particle by a surface plasmon

A. V. Maslov

It is predicted that the optical force induced by a surface plasmon can form a stable equilibrium position for a resonant particle at a finite distance from the surface. The levitated particle can be efficiently propelled along the surface without touching it. The levitation originates from the strong interaction of the particle with the surface.


The effect of saliva on the fate of nanoparticles

Birgit J. Teubl, Biljana Stojkovic, Dominic Docter, Elisabeth Pritz, Gerd Leitinger, Igor Poberaj, Ruth Prassl, Roland H. Stauber, Eleonore Fröhlich, Johannes G. Khinast, Eva Roblegg

The design of nanocarriers for local drug administration to the lining mucosa requires a sound knowledge of how nanoparticles (NPs) interact with saliva. This contact determines whether NPs agglomerate and become immobile due to size- and interaction-filtering effects or adsorb on the cell surface and are internalized by epithelial cells. The aim of this study was to examine the behavior of NPs in saliva considering physicochemical NP properties. The salivary pore–size distribution was determined, and the viscosity of the fluid inside of the pores was studied with optical tweezers. Distinct functionalized NPs (20 and 200 nm) were dispersed in saliva and salivary buffers and characterized, and surface-bound MUC5B and MUC7 were analyzed by 1D electrophoresis and immunoblotting. NP mobility was recorded, and cellular uptake studies were performed with TR146 cells. The mode diameter of the salivary mesh pores is 0.7 μm with a peak width of 1.9 μm, and pores are filled with a low-viscosity fluid. The physicochemical properties of the NPs affected the colloidal stability and mobility: compared with non-functionalized particles, which did not agglomerate and showed a cellular uptake rate of 2.8%, functionalized particles were immobilized, which was correlated with agglomeration and increased binding to mucins. The present study showed that the salivary microstructure facilitates NP adsorption. However, NP size and surface functionalization determine the colloidal stability and cellular interactions.


Equilibrium and out-of-equilibrium mechanics of living mammalian cytoplasm

Gupta, Satish Kumar; Guo, Ming

Living cells are intrinsically non-equilibrium systems. They are driven out of equilibrium by the activity of the molecular motors and other enzymatic processes. This activity along with the ever present thermal agitation results in intracellular fluctuations inside the cytoplasm. In analogy to Brownian motion, the material property of the cytoplasm also influences the characteristics of these fluctuations. In this paper, through a combination of experimentation and theoretical analysis, we show that intracellular fluctuations are indeed due to non-thermal forces at relatively long time-scales, however, are dominated solely by thermal forces at relatively short time-scales. Thus, the cytoplasm of living mammalian cells behaves as an equilibrium material at short time-scales. The mean square displacement of these intracellular fluctuations scales inversely with the cytoplasmic shear modulus in this short time-scale equilibrium regime, and is inversely proportional to the square of the cytoplasmic shear modulus in the long time-scale out-of-equilibrium regime. Furthermore, we deploy passive microrheology based on these fluctuations to extract the mechanical property of the cytoplasm at the high-frequency regime. We show that the cytoplasm of living mammalian cells is a weak elastic gel in this regime; this is in an excellent agreement with an independent micromechanical measurement using optical tweezers.


Laser-Printing and 3D Optical-Control of Untethered Microrobots

Ebubekir Avci, Maria Grammatikopoulou, Guang-Zhong Yang

The two-photon photo-polymerization (2PP) method is used to manufacture an articulated micro-robot for indirect manipulation of cellular structures under laser light. To tackle the stickiness issue between the components of the proposed mechanism, optimizing the contact surface areas is carried out. A step-by-step procedure to manufacture and control an untethered articulated micro-robot under laser light is demonstrated. The manufacture and optical control of a floating multi-component micro-mechanism has not been achieved before. This is significant because it is anticipated that the articulated microrobots could be used in complex biomedical applications where control in 3D space is required such as single-cell analysis, embryo injection, polar-body biopsy, nuclear transplantation, and multi-dimensional imaging for microsurgery.