Discrete dipole approximation for time-domain computation of optical forces on magnetodielectric scatterers

Patrick C. Chaumet, Kamal Belkebir, and Adel Rahmani

We present a general approach, based on the discrete dipole approximation (DDA), for the computation of the exchange of momentum between light and a magnetodielectric, three-dimensional object with arbitrary geometry and linear permittivity and permeability tensors in time domain. The method can handle objects with an arbitrary shape, including objects with dispersive dielectric and/or magnetic material responses.

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Combined Simulation and Experimental Study of Large Deformation of Red Blood Cells in Microfluidic Systems

David J. Quinn, Igor Pivkin, Sophie Y. Wong, Keng-Hwee Chiam, Ming Dao, George Em Karniadakis andSubra Suresh
We investigate the biophysical characteristics of healthy human red blood cells (RBCs) traversing microfluidic channels with cross-sectional areas as small as 2.7 × 3 μm. We combine single RBC optical tweezers and flow experiments with corresponding simulations based on dissipative particle dynamics (DPD), and upon validation of the DPD model, predictive simulations and companion experiments are performed in order to quantify cell deformation and pressure–velocity relationships for different channel sizes and physiologically relevant temperatures. We discuss conditions associated with the shape transitions of RBCs along with the relative effects of membrane and cytosol viscosity, plasma environments, and geometry on flow through microfluidic systems at physiological temperatures. In particular, we identify a cross-sectional area threshold below which the RBC membrane properties begin to dominate its flow behavior at room temperature; at physiological temperatures this effect is less profound.

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Bidirectional Power Stroke by Ncd Kinesin

Anthony E. Butterfield, Russell J. Stewart, Christoph F. Schmidt and Mikhail Skliar

Optical trapping experiments reveal details of molecular motor dynamics. In noisy data, temporal structure within the power stroke of motors can be analyzed by ensemble averaging, but this obscures infrequent subcategories of events. We have here developed an analysis method that uses Kalman filtering of measurements, model-based estimation of the power strokes produced by the motor head, and automatic event classification to discriminate between different types of motor events. This method was applied to optical trap measurements of power strokes of the Drosophila kinesin-14 ncd in a three-bead geometry. We found the majority of events to be consistent with the previously discovered minus-end directed power stroke of ncd, occurring with ATP binding. Unexpectedly, 30% of apparent power strokes were plus-directed and 6% of binding events did not terminate in a discernible stroke. Ensemble averaging for each event category revealed that plus- and minus-directed strokes have different size and occur at different instants within the ncd-MT attachment sequence.

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Brownian Motion of Graphene

Onofrio M. Maragó, Francesco Bonaccorso, Rosalba Saija, Giulia Privitera, Pietro G. Gucciardi, Maria Antonia Iati, Giuseppe Calogero, Philip H. Jones, Ferdinando Borghese, Paolo Denti, Valeria Nicolosi, and Andrea C. Ferrari
Brownian motion is a manifestation of the fluctuation−dissipation theorem of statistical mechanics. It regulates systems in physics, biology, chemistry, and finance. We use graphene as prototype material to unravel the consequences of the fluctuation−dissipation theorem in two dimensions, by studying the Brownian motion of optically trapped graphene flakes. These orient orthogonal to the light polarization, due to the optical constants anisotropy. We explain the flake dynamics in the optical trap and measure force and torque constants from the correlation functions of the tracking signals, as well as comparing experiments with a full electromagnetic theory of optical trapping. The understanding of optical trapping of two-dimensional nanostructures gained through our Brownian motion analysis paves the way to light-controlled manipulation and all-optical sorting of biological membranes and anisotropic macromolecules.

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Ligation of complement receptor 1 increases erythrocyte membrane deformability

Aleksandra M. Glodek, Rossen Mirchev, David E. Golan,Joseph A. Khoory, Jennie M. Burns, Sergey S. Shevkoplyas, Anne Nicholson-Weller and Ionita C. Ghiran
Microbes as well as immune complexes and other continuouslygenerated inflammatory particles are efficiently removed from the human circulation by red blood cells (RBCs) through a process called immune-adherence clearance. During this process, RBCs use complement receptor 1 (CR1, CD35) to bind circulating complement-opsonized particles and transfer them to resident macrophages in the liver and spleen for removal. We here show that ligation of RBC CR1 by antibody and complement-opsonized particles induces a transient Ca++ influx that is proportional to the RBC CR1 levels and is inhibited by T1E3 pAb, a specific inhibitor of TRPC1 channels. The CR1-elicited RBC Ca++ influx is accompanied by an increase in RBC membrane deformability that positively correlates with the number of pre-existing CR1 molecules on RBC membranes. Biochemically, ligation of RBC CR1 causes a significant increase in phosphorylation levels of β-spectrin that is inhibited by pre-incubation of RBCs with DMAT, a specific casein kinase II inhibitor. We hypothesize that the CR1-dependent increase in membrane deformability could be relevant for facilitating the transfer of CR1-bound particles from the RBCs to the hepatic and splenic phagocytes.

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Finite-difference analysis of plasmon-induced forces of metal nano-clusters by the Lorentz force formulation

Masafumi Fujii

We analyze light-induced forces on metal nano-spheres by using the three-dimensional finite-difference time-domain method with the Lorentz force formulation. Convergent analysis of the force on metal nano-particle clusters has been achieved by integrating the Lorentz and the Coulomb forces over the volume of the metal particles. Comparison to the Mie theory of radiation pressure on metal spheres under a plane wave illumination has verified rigorously the accuracy of the numerical method. We also analyze separate two metal spheres in close proximity and the results of the induced forces are compared to those in previous publications. The present method allows analysis of forces on various irregular structures; we apply the method to touching metal spheres, forming a simple cluster with a slight deformation at the contact point, to analyze the forces induced by the plasmonic resonance of the clusters. We show that the fundamental resonance modes, which newly appear in an infrared range when spheres are touching, exhibit strong binding forces within the clusters. Based on the numerical analyses we identify the resonance modes and evaluate quantitatively the infrared-induced forces on metal nano-sphere clusters.

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Optimizing light-matter interaction on the biophotonics workstation

Bañas, A., Palima, D., Tauro, S., Glückstad, J.

Researchers overcome the limitation of range of motion in the axial direction for tweezer-based traps that needed high-intensity regions and high numerical-apertures (NA) by using a counter-propagating beam geometry of the BioPhotonics Workstation. The BioPhotonics Workstation featured real-time reconfigurable counter-propagating beam traps. Intensity patterns defining the optical traps were directly mapped into an addressable light-shaping module, minimizing computational overhead. Axial manipulation was achieved by balancing the intensity ratios of the counter-propagating beams. The axial degree of freedom enabled the flipping of planar microstructures and lifting puzzle pieces of reconfigurable microenvironments. The use of low NA objectives also allowed a wide range of axial manipulation and more freedom on the sample containers.

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Full 3D translational and rotational optical control of multiple rod-shaped bacteria

Florian Hörner, Mike Woerdemann, Stephanie Müller, Berenike Maier, Cornelia Denz

The class of rod-shaped bacteria is an important example of non-spherical objects where defined alignment is desired for the observation of intracellular processes or studies of the flagella. However, all available methods for orientational control of rod-shaped bacteria are either limited with respect to the accessible rotational axes or feasible angles or restricted to one single bacterium. In this paper we demonstrate a scheme to orientate rod-shaped bacteria with holographic optical tweezers (HOT) in any direction. While these bacteria have a strong preference to align along the direction of the incident laser beam, our scheme provides for the first time full rotational control of multiple bacteria with respect to any arbitrary axis. In combination with the translational control HOT inherently provide, this enables full control of all three translational and the two important rotational degrees of freedom of multiple rod-shaped bacteria and allows one to arrange them in any desired configuration.

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Managing Hierarchical Supramolecular Organization with Holographic Tweezers

M. Woerdemann, A. Devaux, L. De Cola, and C. Denz

The use of light for controlling objects led to the development of optical tweezers. Now, the applications have expanded into the creation of three-dimensional structures and the control of surface structures.

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Intangible pointlike tracers for liquid-crystal-based microsensors

Etienne Brasselet and Saulius Juodkazis

We propose an optical detection technique for liquid-crystal-based sensors that is based on polarization-resolved tracking of optical singularities and does not rely on standard observation of light-intensity changes caused by modifications of the liquid crystal orientational ordering. It uses a natural two-dimensional network of polarization singularities embedded in the transverse cross section of a probe beam that passes through a liquid crystal sample, in our case, a nematic droplet held in laser tweezers. The identification and spatial evolution of such a topological fingerprint is retrieved from subwavelength polarization-resolved imaging, and the mechanical constraint exerted on the molecular ordering by the trapping beam itself is chosen as the control parameter. By restricting our analysis to one type of point singularity, C points, which correspond to location in space where the polarization azimuth is undefined, we show that polarization singularities appear as intangible pointlike tracers for liquid-crystal-based three-dimensional microsensors. The method has a superresolution potential and can be used to visualize changes at the nanoscale.

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Microlens-array-enabled on-chip optical trapping and sorting

Xing Zhao, Yuyang Sun, Jing Bu, Siwei Zhu, and X.-C. Yuan

An on-chip optical trapping and sorting system composed of a microchamber and a microlens array (MLA) is demonstrated. The MLA focuses the incident light into multiple confocal spots to trap the particles within the microchamber. The SiO2/ZrO2 solgel material is introduced in the fabrication of MLA for its unique optical and chemical characters. Moreover, in order to prove the effectiveness of the system, experimental demonstration of multibeam trapping and locked-in transport of micropolymer particles in the microchamber is implemented. The system may easily be integrated as microfluidic devices, offering a simple and efficient solution for optical trapping and sorting of biological particles in lab-on-a-chip technologies.

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Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria

Joseph E. Reiner, Rani B. Kishore, Barbara C. Levin,Thomas Albanetti, Nicholas Boire, Ashley Knipe, Kristian Helmerson, Koren Holland Deckman

Background: Mitochondrial DNA (mtDNA) genome mutations can lead to energy and respiratory-related disorders like myoclonic epilepsy with ragged red fiber disease (MERRF), mitochondrial myopathy, encephalopathy, lactic acidosis and stroke (MELAS) syndrome, and Leber's hereditary optic neuropathy (LHON). It is not well understood what effect the distribution of mutated mtDNA throughout the mitochondrial matrix has on the development of mitochondrial-based disorders. Insight into this complex sub-cellular heterogeneity may further our understanding of the development of mitochondria-related diseases. Methodology: This work describes a method for isolating individual mitochondria from single cells and performing molecular analysis on that single mitochondrion's DNA. An optical tweezer extracts a single mitochondrion from a lysed human HL-60 cell. Then a micron-sized femtopipette tip captures the mitochondrion for subsequent analysis. Multiple rounds of conventional DNA amplification and standard sequencing methods enable the detection of a heteroplasmic mixture in the mtDNA from a single mitochondrion. Significance: Molecular analysis of mtDNA from the individually extracted mitochondrion demonstrates that a heteroplasmy is present in single mitochondria at various ratios consistent with the 50/50 heteroplasmy ratio found in single cells that contain multiple mitochondria.

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Deconvolution of dynamic mechanical networks

Michael Hinczewski, Yann von Hansen, and Roland R. Netz

Time-resolved single-molecule biophysical experiments yield data that contain a wealth of dynamic information, in addition to the equilibrium distributions derived from histograms of the time series. In typical force spectroscopic setups the molecule is connected via linkers to a readout device, forming a mechanically coupled dynamic network. Deconvolution of equilibrium distributions, filtering out the influence of the linkers, is a straightforward and common practice. We have developed an analogous dynamic deconvolution theory for the more challenging task of extracting kinetic properties of individual components in networks of arbitrary complexity and topology. Our method determines the intrinsic linear response functions of a given object in the network, describing the power spectrum of conformational fluctuations. The practicality of our approach is demonstrated for the particular case of a protein linked via DNA handles to two optically trapped beads at constant stretching force, which we mimic through Brownian dynamics simulations. Each well in the protein free energy landscape (corresponding to folded, unfolded, or possibly intermediate states) will have its own characteristic equilibrium fluctuations. The associated linear response function is rich in physical content, because it depends both on the shape of the well and its diffusivity—a measure of the internal friction arising from such processes as the transient breaking and reformation of bonds in the protein structure. Starting from the autocorrelation functions of the equilibrium bead fluctuations measured in this force clamp setup, we show how an experimentalist can accurately extract the state-dependent protein diffusivity using a straightforward two-step procedure.

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A novel and effective separation method for single mitochondria analysis

René Pflugradt, Ulrike Schmidt, Benjamin Landenberger, Timo Sänger and Sabine Lutz-Bonengel

To investigate the set of mtDNA molecules contained in small biological structures, powerful techniques for separation are required. We tested flow cytometry (FCM1), laser capture microdissection (LCM2) and a method using optical tweezers (OT3) in combination with a 1μ-Ibidi-Slide with regard to their ability to deposit single mitochondrial particles. The success of separation was determined by real-time quantitative PCR (qPCR4) and sequencing analysis.
OT revealed the highest potential for the separation and deposition of single mitochondrial particles. The study presents a novel setup for effective separation of single mitochondrial particles, which is crucial for the analysis of single mitochondria.

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Angle-suppressed scattering and optical forces on submicrometer dielectric particles

M. Nieto-Vesperinas, R. Gomez-Medina, and J. J. Saenz

We show that submicrometer silicon spheres, whose polarizabilities are completely given by their two first Mie coefficients, are an excellent laboratory to test effects of both angle-suppressed and resonant differential scattering cross sections. Specifically, outstanding scattering angular distributions, with zero forward- or backward-scattered intensity, (i.e., the so-called Kerker conditions), previously discussed for hypothetical magnetodielectric particles, are now observed for those Si objects in the near infrared. Interesting new consequences for the corresponding optical forces are derived from the interplay, both in and out of resonance, between the electric- and magnetic-induced dipoles.

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Optical trapping of spermatozoa using Laguerre-Gaussian laser modes

Raktim Dasgupta, Sunita Ahlawat, Ravi Shanker Verma, Sunita Shukla, and Pradeep Kumar Gupta

We report results of a study on the use of Laguerre-Gaussian (LG) modes for optical trapping of spermatozoa. The results show that for a given trap beam power the first-order LG mode (LG01) leads to lower photodamage to the cells without compromising the trapping efficiency.

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Real-time nonlinear correction of back-focal-plane detection in optical tweezers

Tanuj Aggarwal and Murti Salapaka

Photodiode based detection of laser trapped beads using forward scattered light is a frequently employed technique for position measurement. There is a nonlinear relationship between photodiode outputs and bead position but for small displacements linear approximation holds well. Traditionally, the nonlinearity is compensated by normalizing the photodiode's position signal with the intensity signal and then using a polynomial fit in the range where voltage to position mapping is one to one. In this article, this range is extended by using the intensity signal as an independent input along with the two position signals. A map from the input signals to the actual position values is obtained. This mapping is one-to-one for a larger range that results in an increased detection range. An artificial neural network that facilitates implementation is employed for this purpose. This scheme is implemented on a Field Programmable Gate Array based data acquisition and control hardware with closed loop bandwidth of 50 kHz. Detection of the order of 350 nm from the center of detection laser is demonstrated for a 500 nm diameter bead compared to 180 nm achieved by a polynomial fit.

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Direct Comparison of the Hygroscopic Properties of Ammonium Sulfate and Sodium Chloride Aerosol at Relative Humidities Approaching Saturation

Jim S. Walker, Jon B. Wills, Jonathan P. Reid, Liangyu Wang, David O. Topping, Jason R. Butler, and Yun-Hong Zhang

Holographic optical tweezers are used to make comparative measurements of the hygroscopic properties of single component aqueous aerosol containing sodium chloride and ammonium sulfate over a range of relative humidity from 84% to 96%. The change in RH over the course of the experiment is monitored precisely using a sodium chloride probe droplet with accuracy better than ±0.09%. The measurements are used to assess the accuracy of thermodynamic treatments of the relationship between water activity and solute mass fraction with particular attention focused on the dilute solute limit approaching saturation vapor pressure. The consistency of the frequently used Clegg−Brimblecombe−Wexler (CBW) treatment for predicting the hygroscopic properties of sodium chloride and ammonium sulfate aerosol is confirmed. Measurements of the equilibrium size of ammonium sulfate aerosol are found to agree with predictions to within an uncertainty of ±0.2%. Given the accuracy of treating equilibrium composition, the inconsistencies highlighted in recent calibration measurements of critical supersaturations of sodium chloride and ammonium sulfate aerosol cannot be attributed to uncertainties associated with the thermodynamic predictions and must have an alternative origin. It is concluded that the CBW treatment can allow the critical supersaturation to be estimated for sodium chloride and ammonium sulfate aerosol with an accuracy of better than ±0.002% in RH. This corresponds to an uncertainty of ≤1% in the critical supersaturation for typical supersaturations of 0.2% and above. This supports the view that these systems can be used to accurately calibrate instruments that measure cloud condensation nuclei concentrations at selected supersaturations. These measurements represent the first study in which the equilibrium properties of two particles of chemically distinct composition have been compared simultaneously and directly alongside each other in the same environment.

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Axisymmetric Optical-Trap Measurement of Red Blood Cell Membrane Elasticity

Alexandre Lewalle and Kim H. Parker

The elastic properties of the cell membrane play a crucial role in determining the equilibrium shape of the cell, as well as its response to the external forces it experiences in its physiological environment. Red blood cells are a favored system for studying membrane properties because of their simple structure: a lipid bilayer coupled to a membrane cytoskeleton and no cytoplasmic cytoskeleton. An optical trap is used to stretch a red blood cell, fixed to a glass surface, along its symmetry axis by pulling on a micron-sized latex bead that is bound at the center of the exposed cell dimple. Thesystem, at equilibrium, shows Hookean behavior with a spring constant of 1.5×10−6 N/m over a 1–2 µm range of extension. This choice of simple experimental geometry preserves the axial symmetry of the native cell throughout the stretch, probes membrane deformations in the small-extension regime, and facilitates theoretical analysis. The axisymmetry makes the experiment amenable to simulation using a simple model that makes no a priori assumption on the relative importance of shear and bending in membrane deformations. We use an iterative relaxation algorithm to solve for the geometrical configuration of the membrane at mechanical equilibrium for a range of applied forces. We obtain estimates for the out-of-plane membrane bending modulus B1×10−19 Nm and an upper limit to the in-plane shear modulus H<2×10−6 N/m. The partial agreement of these results with other published values may serve to highlight the dependence of the cell's resistance to deformation on the scale and geometry of the deformation.

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Optical manipulation and binding of microrods with multiple traps enabled in an inclined dual-fiber system

Yuxiang Liu and Miao Yu

We present experimental demonstrations of optical manipulation and optical binding of microscopic glass rods using the multiple traps created by a dual-fiber optical trapping system. Trapping, alignment, rotation, and stacking of glass rods were realized. To the best of our knowledge, this is the first time that cylindrical particles are optically trapped and bound by an optical fiber-based system. The optical manipulation of rods is also investigated through numerical simulations, which are used to quantitatively explain the experimental results. The ability of manipulating multiple particles of different shapes, as well as the integrable nature of the fiber-based setup, bestows the system the potential to be used in microfluidic systems for versatile particle manipulations.

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Optofluidic tweezer on a chip

K. Ono, S. Kaneda, T. Shiraishi, and T. Fujii

A novel method to realize an optical tweezer involving optofluidic operation in a microchannel is proposed. To manipulate the optical tweezer, light from an optical fiber is passed through both PDMS (polydimethylsiloxane)-air surface lenses and an optofluidic region, which is located in a control channel. Two liquids with different refractive indices (RIs) are introduced into the control channel to form two different flow patterns (i.e., laminar and segmented flows), depending on the liquid compositions, the channel geometry, and the flow rates. By altering the shapes of the interface of the two liquids in the optofluidic region, we can continuously or intermittently control the optical paths of the light. To demonstrate the functionality of the proposed method, optical tweezer operations on a chip are performed. Changing the flow pattern of two liquids with different RIs in the optofluidic region results in successful trapping of a 25 μm diameter microsphere and its displacement by 15 μm.

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Optical switching based on the manipulation of microparticles in a colloidal liquid using strong scattering force

Liu Jin, Liu Zheng-Qi, Feng Tian-Hua, Dai Qiao-Feng, Wu Li-Jun and Lan Sheng

This paper demonstrates the realization of an optical switch by optically manipulating a large number of polystyrene spheres contained in a capillary. The strong scattering force exerted on polystyrene spheres with a large diameter of 4.3 μm is employed to realize the switching operation. A transparent window is opened for the signal light when the polystyrene spheres originally located at the beam centre are driven out of the beam region by the strong scattering force induced by the control light. The switching dynamics under different incident powers is investigated and compared with that observed in the optical switch based on the formation of optical matter. It is found that a large extinction ratio of ~ 30 dB and fast switching-on and switching-off times can be achieved in this type of switch.

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Optical twists in phase and amplitude

Vincent R. Daria, Darwin Z. Palima, and Jesper Glückstad

Light beams with helical phase profile correspond to photons having orbital angular momentum (OAM). A Laguerre-Gaussian (LG) beam is an example where its helical phase sets a phase-singularity at the optical axis and forms a ring-shaped transverse amplitude profile. Here, we describe a unique beam where both phase and amplitude express a helical profile as the beam propagates in free space. Such a beam can be accurately referred to as an optical twister. We characterize optical twisters and demonstrate their capacity to induce spiral motion on particles trapped along the twisters’ path. Unlike LG beams, the far field projection of the twisted optical beam maintains a high photon concentration even at higher values of topological charge. Optical twisters have therefore profound applications to fundamental studies of light and atoms such as in quantum entanglement of the OAM, toroidal traps for cold atoms and for optical manipulation of microscopic particles.

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Laser Printing Single Gold Nanoparticles

Alexander S. Urban, Andrey A. Lutich, Fenando D. Stefani, and Jochen Feldmann

Current colloidal synthesis is able to produce an extensive spectrum of nanoparticles with unique optoelectronic, magnetic, and catalytic properties. In order to exploit them in nanoscale devices, flexible methods are needed for the controlled integration of nanoparticles on surfaces with few-nanometer precision. Current technologies usually involve a combination of molecular self-assembly with surface patterning by diverse lithographic methods like UV, dip-pen, or microcontact printing.1,2 Here we demonstrate the direct laser printing of individual colloidal nanoparticles by using optical forces for positioning and the van der Waals attraction for binding them to the substrate. As a proof-of-concept, we print single spherical gold nanoparticles with a positioning precision of 50 nm. By analyzing the printing mechanism, we identify the key physical parameters controlling the method, which has the potential for the production of nanoscale devices and circuits with distinct nanoparticles.

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Multiple Optical Traps with a Single-Beam Optical Tweezer Utilizing Surface Micromachined Planar Curved Grating

Kuo, Ju-Nan; Chen, Kuan-Yu 

In this paper, we present a single-beam optical tweezer integrated with a planar curved diffraction grating for microbead manipulation. Various curvatures of the surface micromachined planar curved grating are systematically investigated. The planar curved grating was fabricated using multiuser micro-electro-mechanical-system (MEMS) processes (MUMPs). The angular separation and the number of diffracted orders were determined. Experimental results indicate that the diffraction patterns and curvature of the planar curved grating are closely related. As the curvature of the planar curved grating increases, the vertical diffraction angle increases, resulting in the strip patterns of the planar curved grating. A single-beam optical tweezer integrated with a planar curved diffraction grating was developed. We demonstrate a technique for creating multiple optical traps from a single laser beam using the developed planar curved grating. The strip patterns of the planar curved grating that resulted from diffraction were used to trap one row of polystyrene beads.

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A new method for the covalent attachment of DNA to a surface for single-molecule studies

Daniel J. Schlingman, Andrew H. Mack, Simon G.J. Mochrie and Lynne Regan

Attachments between DNA and a surface or bead are often necessary for single-molecule studies of DNA and DNA–protein interactions. In single-molecule mechanical studies using optical or magnetic tweezers, such attachments must be able to withstand the applied forces. Here we present a new method for covalently attaching DNA to a glass surface, which uses N-hydroxysuccinimide (NHS) modified PEG that is suitable for high-force single-molecule mechanical studies. A glass surface is coated with silane-PEG-NHS and DNA is covalently linked through a reaction between the NHS group and an amine modified nucleotide that has been incorporated into the DNA. After DNA attachment, non-reacted NHS groups are hydrolyzed leaving a PEG-covered surface which has the added benefit of reducing non-specific surface interactions. This method permits specific binding of the DNA to the surface through a covalent bond. At the DNA end not attached to the surface, we attach a streptavidin-coated polystyrene bead and measure force-versus-extension using an optical trap. We show that our method allows a tethered DNA molecule to be pulled through its overstretching transition (>60 pN) multiple times. We anticipate this simple yet powerful method will be useful for many researchers.

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Monitoring and rapid quantification of total carotenoids in Rhodotorula glutinis cells using laser tweezers Raman spectroscopy

Zhanhua Tao, Guiwen Wang, Xiaodong Xu, Yufeng Yuan, Xue Wang, Yongqing Li

Rhodotorula glutinis is known to accumulate large amounts of carotenoids under certain culture conditions, which have very important industrial applications. So far, the molecular mechanism of regulating carotenogenesis is still not well understood. To better understand the carotenogenesis process, it requires methods that can detect carotenogenesis rapidly and reliably in single live cells. In this paper, a method based on laser tweezers Raman spectroscopy (LTRS) was developed to directly detect carotenoids, as well as other important biological molecules in single live R. glutinis cells. The data showed that the accumulation of carotenoids and lipids occurred mainly in the late exponential and stationary phases when the cell growth was inhibited by nutrient limitation. Meanwhile, the carotenoid concentration changed together with the concentration of nucleic acids, which increased in the first phase and decreased in the last phase of the culture. These data demonstrate that LTRS is a rapid, convenient, and reliable method to study the carotenogenesis processin vivo.

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Mapping the classical cross-bridge theory and backward steps in a three bead laser trap setup

G. Schappacher-Tilp, A. Jinha and W. Herzog

According to the cross-bridge theory (Huxley, 1957) [1], the interaction between myosin and actin is governed by a deterministic process where the myosin molecule pulls the actin filament in one specific direction only. However, studies on single myosin–actin interactions produced displacements of actin not only in the preferred but also in the opposite direction. This phenomenon is typically referred to as backward steps by the myosin head. Molloy et al. (1995)[2] speculated that these backward steps are not caused by the molecular interactions of actin with myosin but are an artifact of the Brownian motion associated with these molecular level experiments. The aim of this study was to investigate, whether a theoretical model can support Molloy’s speculation. We therefore developed a theoretical model of actin–myosin based muscle contraction that was strictly based on Huxley’s assumption of one stepping direction only, but incorporated Brownian motion, as observed in single cross-bridge-actin interactions. The mathematical model is based on Langevin equations describing the classical three-bead laser trap setup and uses a novel semi-analytical approach to study the percentage of backward steps. We analyzed the effects of different initial actin attachment site distribution and laser trap stiffness on the ratio of forward to backward steps. Our results demonstrate that backward steps and the classical cross-bridge theory are perfectly compatible in a three-bead laser trap setup.

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Spatial light modulators for the manipulation of individual atoms

L. Brandt, C. Muldoon, T. Thiele, J. Dong, E. Brainis and A. Kuhn

We propose a versatile arrangement for the trapping and manipulation of single atoms in optical tweezers formed by the direct image of a spatial light modulator (SLM). The scheme incorporates a high numerical aperture microscope to map the intensity distribution of a SLM onto a cloud of cold atoms. The regions of high intensity act as optical dipole-force traps. With a SLM fast enough to modify the trapping potential in real time, this technique is well suited for the controlled addressing and manipulation of arbitrarily selected atoms.

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Single-Molecule Adhesion Forces and Attachment Lifetimes of Myosin-I Phosphoinositide Interactions

Serapion Pyrpassopoulos, Henry Shuman and E. Michael Ostap

Phosphoinositides regulate the activities and localization of many cytoskeletal proteins involved in crucial biological processes, including membrane-cytoskeleton adhesion. Yet little is known about the mechanics of protein-phosphoinositide interactions, or about the membrane-attachment mechanics of any peripheral membrane proteins. Myosin-Ic (myo1c) is a molecular motor that links membranes to the cytoskeleton via phosphoinositide binding, so it is particularly important to understand the mechanics of its membrane attachment. We used optical tweezers to measure the strength and attachment lifetime of single myo1c molecules as they bind beads coated with a bilayer of 2% phosphatidylinositol 4,5-bisphosphate and 98% phosphatidylcholine. Adhesion forces measured under ramp-load ranged between 5.5 and 16 pN at loading rates between 250 and 1800 pN/s. Dissociation rates increased linearly with constant force (0.3–2.5 pN), with rates exceeding 360 s−1 at 2.5 pN. Attachment lifetimes calculated from adhesion force measurements were loading-rate-dependent, suggesting nonadiabatic behavior during pulling. The adhesion forces of myo1c with phosphoinositides are greater than the motors stall forces and are within twofold of the force required to extract a lipid molecule from the membrane. However, attachment durations are short-lived, suggesting that phosphoinositides alone do not provide the mechanical stability required to anchor myo1c to membranes during multiple ATPase cycles.

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Published Papers Statistics for 2010

Here is the long awaited results for the year of 2010 for published papers on optical tweezers, micromanipulation and trapping, in this blog (a total of 368 posts for 2010).

The top Journals (more than 2% hits) are:

1. Optics Express                         13.5%
2. Optics Letters                             4.4%
3. Physical Review E                      3.6%
4. Physical Review Letters             3.0%
5. Proc. of the Nat. Acad. of Sci.   3.0%
6. Nano Letters                                2.7%
7. Biophysical Journal                     2.5%
8. Journal of Biomedical Optics     2.5%
9. Physical Review A                       2.5%
10. Lab on a Chip                               2.2 %

Below is also a tag cloud (from the words found in the title and abstracts for 2010):