We unzip DNA molecules using optical tweezers and determine the sizes of the cooperatively unzipping and zipping regions separating consecutive metastableintermediates along the unzipping pathway. Sizes are found to be distributed following a power law, ranging from one base pair up to more than a hundred base pairs. We find that a large fraction of unzipping regions smaller than 10 bp are seldom detected because of the high compliance of the released single stranded DNA. We show how the compliance of a single nucleotide sets a limit value around 0.1 N/m for the stiffness of any local force probe aiming to discriminate one base pair at a time in DNA unzipping experiments.
Thursday, December 24, 2009
Statistical Properties of Metastable Intermediates in DNA Unzipping
J. M. Huguet, N. Forns, and F. Ritort
We unzip DNA molecules using optical tweezers and determine the sizes of the cooperatively unzipping and zipping regions separating consecutive metastableintermediates along the unzipping pathway. Sizes are found to be distributed following a power law, ranging from one base pair up to more than a hundred base pairs. We find that a large fraction of unzipping regions smaller than 10 bp are seldom detected because of the high compliance of the released single stranded DNA. We show how the compliance of a single nucleotide sets a limit value around 0.1 N/m for the stiffness of any local force probe aiming to discriminate one base pair at a time in DNA unzipping experiments.
We unzip DNA molecules using optical tweezers and determine the sizes of the cooperatively unzipping and zipping regions separating consecutive metastableintermediates along the unzipping pathway. Sizes are found to be distributed following a power law, ranging from one base pair up to more than a hundred base pairs. We find that a large fraction of unzipping regions smaller than 10 bp are seldom detected because of the high compliance of the released single stranded DNA. We show how the compliance of a single nucleotide sets a limit value around 0.1 N/m for the stiffness of any local force probe aiming to discriminate one base pair at a time in DNA unzipping experiments.
Wednesday, December 23, 2009
Laser Manipulation by Using Liquid Crystal Devices with Variable-Focusing and Beam-Steering Functions
Marenori Kawamura; Junji Onishi; Susumu Sato
A laser manipulation system for trapping and controlling the positions of microscopic transparent particles by using liquid crystal (LC) devices is developed. The LC device has functions of variable-focusing and beam-steering by controlling the applied voltages to the LC device without mechanical movements. The trapped particles suspended in deionized water can easily be shifted along the position of the focused laser spot. The microscopic rod-like particles can also be shifted and rotated in the clockwise or anticlockwise along the direction of the major axis of the elliptically distributed beam intensity. In addition, the multiple microscopic particles at the bright region of the linear interference fringe patterns of the LC device with comb-shaped electrodes can be trapped and shifted along the fringe patterns.
A laser manipulation system for trapping and controlling the positions of microscopic transparent particles by using liquid crystal (LC) devices is developed. The LC device has functions of variable-focusing and beam-steering by controlling the applied voltages to the LC device without mechanical movements. The trapped particles suspended in deionized water can easily be shifted along the position of the focused laser spot. The microscopic rod-like particles can also be shifted and rotated in the clockwise or anticlockwise along the direction of the major axis of the elliptically distributed beam intensity. In addition, the multiple microscopic particles at the bright region of the linear interference fringe patterns of the LC device with comb-shaped electrodes can be trapped and shifted along the fringe patterns.
Monday, December 21, 2009
Trapping of low-refractive-index particles with azimuthally polarized beam
Fei Peng, Baoli Yao, Shaohui Yan, Wei Zhao, and Ming Lei
Azimuthally polarized beams, focused by a high-numerical-aperture (NA) objective lens, form a hollow intensity distribution near the focus, which is appropriate for trapping low-refractive-index particles, in contrast to common linearly polarized or radially polarized beams. In this paper, the field distribution of the azimuthally polarized beam focused by a high-NA objective is described by the vectorial diffraction integral, and then the radiation forces on spherical particles with different parameters such as radius and refractive index are calculated by the T-matrix method. Numerical results show that the azimuthally polarized beam not only can steadily trap low-refractive-index particles at the focus center but also can trap multiple high-refractive-index particles around the focus center by virtue of the hollow-ring configuration. The range of the sizes of low-refractive-index particles that can be trapped steadily are presented, corresponding to different parameters such as the NA of the objective and the relative refractive index, based on which the NA of the objective can be selected to trap the appropriate size of particles.
Azimuthally polarized beams, focused by a high-numerical-aperture (NA) objective lens, form a hollow intensity distribution near the focus, which is appropriate for trapping low-refractive-index particles, in contrast to common linearly polarized or radially polarized beams. In this paper, the field distribution of the azimuthally polarized beam focused by a high-NA objective is described by the vectorial diffraction integral, and then the radiation forces on spherical particles with different parameters such as radius and refractive index are calculated by the T-matrix method. Numerical results show that the azimuthally polarized beam not only can steadily trap low-refractive-index particles at the focus center but also can trap multiple high-refractive-index particles around the focus center by virtue of the hollow-ring configuration. The range of the sizes of low-refractive-index particles that can be trapped steadily are presented, corresponding to different parameters such as the NA of the objective and the relative refractive index, based on which the NA of the objective can be selected to trap the appropriate size of particles.
Multi-mode mitigation in an optofluidic chip for particle manipulation and sensing
Philip Measor, Sergei Kühn, Evan J. Lunt, Brian S. Phillips, Aaron R. Hawkins, andHolger Schmidt
A new waveguide design for an optofluidic chip is presented. It mitigates multi-mode behavior in solid and liquid-core waveguides by increasing fundamental mode coupling to 82% and 95%, respectively. Additionally, we demonstrate a six-fold improvement in lateral confinement of optically guided dielectric microparticles and double the detection efficiency of fluorescent particles.
A new waveguide design for an optofluidic chip is presented. It mitigates multi-mode behavior in solid and liquid-core waveguides by increasing fundamental mode coupling to 82% and 95%, respectively. Additionally, we demonstrate a six-fold improvement in lateral confinement of optically guided dielectric microparticles and double the detection efficiency of fluorescent particles.
Friday, December 18, 2009
An Optical Conveyor for Molecules
Franz M. Weinert and Dieter Braun
Trapping single ions under vacuum allows for precise spectroscopy in atomic physics. The confinement of biological molecules in bulk water is hindered by the lack of comparably strong forces. Molecules have been immobilized to surfaces, however often with detrimental effects on their function. Here, we optically trap molecules by creating the microscale analogue of a conveyor belt: a bidirectional flow is combined with a perpendicular thermophoretic molecule drift. Arranged in a toroidal geometry, the conveyor accumulates a hundredfold excess of 5-base DNA within seconds. The concentrations of the trapped DNA scale exponentially with length, reaching trapping potential depths of 14 kT for 50 bases. The mechanism does not require microfluidics, electrodes, or surface modifications. As a result, the trap can be dynamically relocated. The optical conveyor can be used to enhance diffusion-limited surface reactions, redirect cellular signaling, observe individual biomolecules over a prolonged time, or approach single-molecule chemistry in bulk water.
Trapping single ions under vacuum allows for precise spectroscopy in atomic physics. The confinement of biological molecules in bulk water is hindered by the lack of comparably strong forces. Molecules have been immobilized to surfaces, however often with detrimental effects on their function. Here, we optically trap molecules by creating the microscale analogue of a conveyor belt: a bidirectional flow is combined with a perpendicular thermophoretic molecule drift. Arranged in a toroidal geometry, the conveyor accumulates a hundredfold excess of 5-base DNA within seconds. The concentrations of the trapped DNA scale exponentially with length, reaching trapping potential depths of 14 kT for 50 bases. The mechanism does not require microfluidics, electrodes, or surface modifications. As a result, the trap can be dynamically relocated. The optical conveyor can be used to enhance diffusion-limited surface reactions, redirect cellular signaling, observe individual biomolecules over a prolonged time, or approach single-molecule chemistry in bulk water.
Thursday, December 17, 2009
Dynamic micro-bead arrays using optical tweezers combined with intelligent control techniques
Yoshio Tanaka, Hiroyuki Kawada, Shogo Tsutsui, Mitsuru Ishikawa, and Hiroyuki Kitajima
Dynamic micro-bead arrays offer great flexibility and potential as sensing tools in various scientific fields. Here we present a software-oriented approach for fully automated assembly of versatile dynamic micro-bead arrays using multi-beam optical tweezers combined with intelligent control techniques. Four typical examples, including the collision-free sorting of array elements by bead features, are demonstrated in real time. Control algorithms and experimental apparatus for these demonstrations are also described.
Dynamic micro-bead arrays offer great flexibility and potential as sensing tools in various scientific fields. Here we present a software-oriented approach for fully automated assembly of versatile dynamic micro-bead arrays using multi-beam optical tweezers combined with intelligent control techniques. Four typical examples, including the collision-free sorting of array elements by bead features, are demonstrated in real time. Control algorithms and experimental apparatus for these demonstrations are also described.
Spatial Stability of Particles Trapped by Time-Division Optical Tweezers
Johtaro Yamamoto; Toshiaki Iwai
We developed an on-demand multiple-spot holographic optical tweezers (HOT) system based on quasi-simultaneous generation of two intensity-spot patterns by alternately sending the two corresponding hologram patterns to a spatial light modulator. This switching operation reduces the spatial stability of a Brownian particle trapped inside the generated intensity spot. In this study, numerical analysis of the conditions for stable particle trapping in the time-division HOT is conducted using the Smoluchowski equation under the Rayleigh scattering approximation. The relationship between the particle size, the switching rate, and the focused laser beam power is obtained. Experiments confirm the validity of the numerical analysis.
We developed an on-demand multiple-spot holographic optical tweezers (HOT) system based on quasi-simultaneous generation of two intensity-spot patterns by alternately sending the two corresponding hologram patterns to a spatial light modulator. This switching operation reduces the spatial stability of a Brownian particle trapped inside the generated intensity spot. In this study, numerical analysis of the conditions for stable particle trapping in the time-division HOT is conducted using the Smoluchowski equation under the Rayleigh scattering approximation. The relationship between the particle size, the switching rate, and the focused laser beam power is obtained. Experiments confirm the validity of the numerical analysis.
Passive and active microrheology for cross-linked F-actin networks in vitro
Hyungsuk Lee, Jorge M. Ferrer, Fumihiko Nakamura, Matthew J. Lang and Roger D. Kamm
Actin filament (F-actin) is one of the dominant structural constituents in the cytoskeleton. Orchestrated by various actin-binding proteins (ABPs), F-actin is assembled into higher-order structures such as bundles and networks that provide mechanical support for the cell and play important roles in numerous cellular processes. Although mechanical properties of F-actin networks have been extensively studied, the underlying mechanisms for network elasticity are not fully understood, in part because different measurements probe different length and force scales. Here, we developed both passive and active microrheology techniques using optical tweezers to estimate the mechanical properties of F-actin networks at a length scale comparable to cells. For the passive approach we tracked the motion of a thermally fluctuating colloidal sphere to estimate the frequency-dependent complex shear modulus of the network. In the active approach, we used an optical trap to oscillate an embedded microsphere and monitored the response in order to obtain network viscoelasticity over a physiologically relevant force range. While both active and passive measurements exhibit similar results at low strain, the F-actin network subject to high strain exhibits non-linear behavior which is analogous to the strain-hardening observed in macroscale measurements. Using confocal and total internal reflection fluorescent microscopy, we also characterize the microstructure of reconstituted F-actin networks in terms of filament length, mesh size and degree of bundling. Finally, we propose a model of network connectivity by investigating the effect of filament length on the mechanical properties and structure.
Direct Measurement of Shear-Induced Cross-Correlations of Brownian Motion
A. Ziehl, J. Bammert, L. Holzer, C. Wagner, and W. Zimmermann
Shear-induced cross-correlations of particle fluctuations perpendicular and along streamlines are investigated experimentally and theoretically. Direct measurements of the Brownian motion of micron-sized beads, held by optical tweezers in a shear-flow cell, show a strong time asymmetry in the cross-correlation, which is caused by the non-normal amplification of fluctuations. Complementary measurements on the single particle probability distribution substantiate this behavior and both results are consistent with a Langevin model. In addition, a shear-induced anticorrelation between orthogonal random displacements of two trapped and hydrodynamically interacting particles is detected, having one or two extrema in time, depending on the positions of the particles.
Shear-induced cross-correlations of particle fluctuations perpendicular and along streamlines are investigated experimentally and theoretically. Direct measurements of the Brownian motion of micron-sized beads, held by optical tweezers in a shear-flow cell, show a strong time asymmetry in the cross-correlation, which is caused by the non-normal amplification of fluctuations. Complementary measurements on the single particle probability distribution substantiate this behavior and both results are consistent with a Langevin model. In addition, a shear-induced anticorrelation between orthogonal random displacements of two trapped and hydrodynamically interacting particles is detected, having one or two extrema in time, depending on the positions of the particles.
Wednesday, December 16, 2009
Optical tweezer for probing erythrocyte membrane deformability
Manas Khan, Harsh Soni, and A. K. Sood
We report that the average rotation speed of optically trapped crenated erythrocytes is direct signature of their membrane deformability. When placed in hypertonic buffer, discocytic erythrocytes are subjected to crenation. The deformation of cells brings in chirality and asymmetry in shape that makes them rotate under the scattering force of alinearly polarized optical trap. A change in the deformability of the erythrocytes, due to any internal or environmental factor, affects the rotation speed of the trapped crenated cells. Here we show how the increment in erythrocyte membrane rigidity with adsorption of Ca++ions can be exhibited through this approach.
We report that the average rotation speed of optically trapped crenated erythrocytes is direct signature of their membrane deformability. When placed in hypertonic buffer, discocytic erythrocytes are subjected to crenation. The deformation of cells brings in chirality and asymmetry in shape that makes them rotate under the scattering force of alinearly polarized optical trap. A change in the deformability of the erythrocytes, due to any internal or environmental factor, affects the rotation speed of the trapped crenated cells. Here we show how the increment in erythrocyte membrane rigidity with adsorption of Ca++ions can be exhibited through this approach.
Electrophoretic Force on a Protein-Coated DNA Molecule in a Solid-State Nanopore
Adam R. Hall, Stijn van Dorp, Serge G. Lemay and Cees Dekker
Using solid-state nanopores with optical tweezers, we perform force spectroscopy on DNA molecules that are coated with RecA proteins. We observe that the electrophoretic force is 2−4 times larger for RecA-DNA filaments than for uncoated DNA molecules and that this force increases at lower salt concentrations. The data demonstrate the efficacy of solid-state nanopores for locally probing the forces on DNA-bound proteins. Our results are described quantitatively by a model that treats the electrophoretic and hydrodynamic forces. The conductance steps that occur when RecA-DNA enters the nanopore change from conductance decreases at high salt to conductance increases at low salt, which allows the apparent charge of the RecA-DNA filament to be extracted. The combination of conductance measurements with local force spectroscopy increases the potential for future solid-state nanopore screening devices.
Using solid-state nanopores with optical tweezers, we perform force spectroscopy on DNA molecules that are coated with RecA proteins. We observe that the electrophoretic force is 2−4 times larger for RecA-DNA filaments than for uncoated DNA molecules and that this force increases at lower salt concentrations. The data demonstrate the efficacy of solid-state nanopores for locally probing the forces on DNA-bound proteins. Our results are described quantitatively by a model that treats the electrophoretic and hydrodynamic forces. The conductance steps that occur when RecA-DNA enters the nanopore change from conductance decreases at high salt to conductance increases at low salt, which allows the apparent charge of the RecA-DNA filament to be extracted. The combination of conductance measurements with local force spectroscopy increases the potential for future solid-state nanopore screening devices.
Monday, December 14, 2009
Wavelength dependence of optical tweezer trapping forces on dye-doped polystyrene microspheres
M. J. Kendrick, D. H. McIntyre, and O. Ostroverkhova
We present an experimental and numerical study of the wavelength dependence, near resonance, of the optical tweezer trap stiffness on three different dye-doped 1 μm polystyrene spheres with peak absorptions at λ=625, 775, and 840 nm. Experimentally, an increase in the trap stiffness of ~35% on the red side of resonance was observed for the dye-doped spheres relative to polystyrene spheres without dye. Numerical simulations for spheres of different sizes, between 20 nm and 1 μm, and for absorption strengths corresponding to peak extinction coefficient values between 0.0027 and 0.081 were also conducted. Numerical results showed a maximum increase in the trap stiffness of ~35%, which is consistent with experimental results.
We present an experimental and numerical study of the wavelength dependence, near resonance, of the optical tweezer trap stiffness on three different dye-doped 1 μm polystyrene spheres with peak absorptions at λ=625, 775, and 840 nm. Experimentally, an increase in the trap stiffness of ~35% on the red side of resonance was observed for the dye-doped spheres relative to polystyrene spheres without dye. Numerical simulations for spheres of different sizes, between 20 nm and 1 μm, and for absorption strengths corresponding to peak extinction coefficient values between 0.0027 and 0.081 were also conducted. Numerical results showed a maximum increase in the trap stiffness of ~35%, which is consistent with experimental results.
Wednesday, December 9, 2009
Design considerations for micro- and nanopositioning: leveraging the latest for biophysical applications
Jordan SC, Anthony PC.
Biophysical applications ranging from fluorescence microassays to single-molecule microscopy are increasingly dependent on automated nanoscale positional control and stability. A whirlwind of motion-industry innovation has resulted in an array of new motion options offering significant improvements in application performance, reproducibility and throughput. The challenge to leverage these developments depends on researchers, engineers and motion vendors acquiring a common language of specifications and a shared understanding of the challenges posed by application needs. To assist in building this shared understanding, this article reviews today's motion technologies, beginning with a concise review of key principles of motion control focusing on applications. It progresses through illustrations of sensor/encoder technologies and servo techniques. A spectrum of classical and recent motion technologies is explored, from stepper and servo actuation of conventional microscopy stages, to advanced piezo stack nanopositioners capable of picometer precision, to novel ultrasonic resonant piezomotors and piezo-ceramic-based mechanisms capable of high-force positioning over many millimeters while providing resolutions down into the sub-nanometer range. A special emphasis is placed on the effects of integrating multiple motion technologies into an application, such as stacking a fine nanopositioner atop a long-travel stage. Examples and data are presented to clarify these issues, including important and insightful new stability measurements taken directly from an advanced optical trapping application. The important topics of software and interfacing are also explored from an applications perspective, since design-and-debugging time, synchronization capabilities and overall throughput are heavily dependent on these often-overlooked aspects of motion system design. The discussion is designed to illuminate specifications-related topics that become increasingly important as precision requirements tighten. Throughout, both traditional and novel techniques and approaches are explored so that readers are left with a solid overview of the state of the art, and an actionable perspective that readies them to discuss and evaluate specifications and vendor capabilities against practical application requirements.
Biophysical applications ranging from fluorescence microassays to single-molecule microscopy are increasingly dependent on automated nanoscale positional control and stability. A whirlwind of motion-industry innovation has resulted in an array of new motion options offering significant improvements in application performance, reproducibility and throughput. The challenge to leverage these developments depends on researchers, engineers and motion vendors acquiring a common language of specifications and a shared understanding of the challenges posed by application needs. To assist in building this shared understanding, this article reviews today's motion technologies, beginning with a concise review of key principles of motion control focusing on applications. It progresses through illustrations of sensor/encoder technologies and servo techniques. A spectrum of classical and recent motion technologies is explored, from stepper and servo actuation of conventional microscopy stages, to advanced piezo stack nanopositioners capable of picometer precision, to novel ultrasonic resonant piezomotors and piezo-ceramic-based mechanisms capable of high-force positioning over many millimeters while providing resolutions down into the sub-nanometer range. A special emphasis is placed on the effects of integrating multiple motion technologies into an application, such as stacking a fine nanopositioner atop a long-travel stage. Examples and data are presented to clarify these issues, including important and insightful new stability measurements taken directly from an advanced optical trapping application. The important topics of software and interfacing are also explored from an applications perspective, since design-and-debugging time, synchronization capabilities and overall throughput are heavily dependent on these often-overlooked aspects of motion system design. The discussion is designed to illuminate specifications-related topics that become increasingly important as precision requirements tighten. Throughout, both traditional and novel techniques and approaches are explored so that readers are left with a solid overview of the state of the art, and an actionable perspective that readies them to discuss and evaluate specifications and vendor capabilities against practical application requirements.
Effect of antibiotics and antimicrobial peptides on single protein motilit
Winther T, Oddershede LB.
Following the movement of individual molecules of a bacterial surface protein in vivo we investigated the effects of antibiotics and antimicrobial peptides on protein motility and membrane structure. In previous work we engineered the lambda-receptor of Escherichia coli such that less than one receptor per cell is in vivo biotinylated and can bind to a streptavidin coated bead. Such a bead served as a handle for the optical tweezers to follow the motion of an individual receptor. In an un-perturbed living cell the lambda-receptor performs a confined diffusive motion. The lambda-receptor links to the peptidoglycan layer, and indeed, a perturbation of the peptidoglycan layer had a pronounced effect on the motility of the receptor: The motility significantly decreases upon treatment with vancomycin or ampicillin, to study the effect of vancomycin we used strains with increased membrane permeability. As the motility of an individual receptor was monitored over an extended amount of time we were able to observe a temporal evolution of the action of vancomycin. Antimicrobial peptides (AMPs) are alternatives to conventional antibiotics in the treatment of bacterial infections. Therefore, we also investigated the effect of the toxic AMP polymyxin B (PMB) which targets both the outer and inner membranes and kills the organism. PMB significantly decreased the motility of the lambda-receptor. On the basis of these findings we confirm that the lambda-receptor is firmly attached to the peptidoglycan layer, and that an antibiotic or AMP mediated destruction of the dynamic peptidoglycan synthesis decreases the receptor motion.
Following the movement of individual molecules of a bacterial surface protein in vivo we investigated the effects of antibiotics and antimicrobial peptides on protein motility and membrane structure. In previous work we engineered the lambda-receptor of Escherichia coli such that less than one receptor per cell is in vivo biotinylated and can bind to a streptavidin coated bead. Such a bead served as a handle for the optical tweezers to follow the motion of an individual receptor. In an un-perturbed living cell the lambda-receptor performs a confined diffusive motion. The lambda-receptor links to the peptidoglycan layer, and indeed, a perturbation of the peptidoglycan layer had a pronounced effect on the motility of the receptor: The motility significantly decreases upon treatment with vancomycin or ampicillin, to study the effect of vancomycin we used strains with increased membrane permeability. As the motility of an individual receptor was monitored over an extended amount of time we were able to observe a temporal evolution of the action of vancomycin. Antimicrobial peptides (AMPs) are alternatives to conventional antibiotics in the treatment of bacterial infections. Therefore, we also investigated the effect of the toxic AMP polymyxin B (PMB) which targets both the outer and inner membranes and kills the organism. PMB significantly decreased the motility of the lambda-receptor. On the basis of these findings we confirm that the lambda-receptor is firmly attached to the peptidoglycan layer, and that an antibiotic or AMP mediated destruction of the dynamic peptidoglycan synthesis decreases the receptor motion.
10 Years of Tension on Chromatin: Results from Single Molecule Force Spectroscopy
Chien, Fan-Tso; van Noort, John
The compact, yet dynamic organization of chromatin plays an essential role in regulating gene expression. Although the static structure of chromatin fibers has been studied extensively, the controversy about the higher order folding remains. In the past ten years a number of studies have addressed chromatin folding with single molecule force spectroscopy. By manipulating chromatin fibers individually, the mechanical properties of the fibers were quantified with piconewton and nanometer accuracy. Here, we review the results of force induced chromatin unfolding and compare the differences between experimental conditions and single molecule manipulation techniques like force and position clamps. From these studies, five major features appeared upon forced extension of chromatin fibers: the elastic stretching of chromatin's higher order structure, the breaking of internucleosomal contacts, unwrapping of the first turn of DNA, unwrapping of the second turn of DNA, and the dissociation of histone octamers. These events occur sequentially at the increasing force. Resolving force induced structural changes of chromatin fibers at the single molecule level will help to provide a physical understanding of processes involving chromatin that occur in vivo and will reveal the mechanical constraints that are relevant for processing and maintenance of DNA in eukaryotes.
The compact, yet dynamic organization of chromatin plays an essential role in regulating gene expression. Although the static structure of chromatin fibers has been studied extensively, the controversy about the higher order folding remains. In the past ten years a number of studies have addressed chromatin folding with single molecule force spectroscopy. By manipulating chromatin fibers individually, the mechanical properties of the fibers were quantified with piconewton and nanometer accuracy. Here, we review the results of force induced chromatin unfolding and compare the differences between experimental conditions and single molecule manipulation techniques like force and position clamps. From these studies, five major features appeared upon forced extension of chromatin fibers: the elastic stretching of chromatin's higher order structure, the breaking of internucleosomal contacts, unwrapping of the first turn of DNA, unwrapping of the second turn of DNA, and the dissociation of histone octamers. These events occur sequentially at the increasing force. Resolving force induced structural changes of chromatin fibers at the single molecule level will help to provide a physical understanding of processes involving chromatin that occur in vivo and will reveal the mechanical constraints that are relevant for processing and maintenance of DNA in eukaryotes.
Single-Molecule Force Spectroscopy Using the NanoTracker™ Optical Tweezers Platform: from Design to Application
Wozniak, A.; van Mameren, J.; Ragona, S.
Since the development of detection and analysis techniques for optical tweezers setups, there has been an everincreasing interest in optical tweezers as a quantitative method, shifting its applications from a pure manipulation tool towards the investigation of motions and forces. With the capability of manipulation and detection of forces of a few hundred picoNewtons down to a fraction of a picoNewton, optical tweezers are perfectly suitable for the investigation of single molecules. Accordingly, the technique has been extensively used for the biophysical characterization of biomolecules, ranging from the mechanical and elastic properties of biological polymers to the dynamics associated with enzymatic activity and protein motility. Here, the use of state-of-the-art optical tweezers on the elasticity of single DNA molecules is presented, highlighting the possibilities this technique offers for the investigation of protein-DNA interaction, but also for other single molecule applications. Technical in nature, design aspects of the NanoTracker™ optical tweezers setup are addressed, presenting the recent advances in the development of optical tweezers, ranging from noise reduction to detection and calibration methodology.
DOI
Since the development of detection and analysis techniques for optical tweezers setups, there has been an everincreasing interest in optical tweezers as a quantitative method, shifting its applications from a pure manipulation tool towards the investigation of motions and forces. With the capability of manipulation and detection of forces of a few hundred picoNewtons down to a fraction of a picoNewton, optical tweezers are perfectly suitable for the investigation of single molecules. Accordingly, the technique has been extensively used for the biophysical characterization of biomolecules, ranging from the mechanical and elastic properties of biological polymers to the dynamics associated with enzymatic activity and protein motility. Here, the use of state-of-the-art optical tweezers on the elasticity of single DNA molecules is presented, highlighting the possibilities this technique offers for the investigation of protein-DNA interaction, but also for other single molecule applications. Technical in nature, design aspects of the NanoTracker™ optical tweezers setup are addressed, presenting the recent advances in the development of optical tweezers, ranging from noise reduction to detection and calibration methodology.
DOI
Transportin Mediates Nuclear Entry of DNA in Vertebrate Systems
Aurelie Lachish-Zalait, Corine K. Lau, Boris Fichtman, Ella Zimmerman, Amnon Harel, Michelle R. Gaylord , Douglass J. Forbes and Michael Elbaum
Delivery of DNA to the cell nucleus is an essential step in many types of viral infection, transfection, gene transfer by the plant pathogen Agrobacteriumtumefaciens and in strategies for gene therapy. Thus, the mechanism by which DNA crosses the nuclear pore complex (NPC) is of great interest. Using nuclei reconstituted in vitro inXenopus egg extracts, we previously studied DNA passage through the nuclear pores using a single-molecule approach based on optical tweezers. Fluorescently labeled DNA molecules were also seen to accumulate within nuclei. Here we find that this import of DNA relies on a soluble protein receptor of the importin family. To identify this receptor, we used different pathway-specific cargoes in competition studies as well as pathway-specific dominant negative inhibitors derived from the nucleoporin Nup153. We found that inhibition of the receptor transportin suppresses DNA import. In contrast, inhibition of importin β has little effect on the nuclear accumulation of DNA. The dependence on transportin was fully confirmed in assays using permeabilized HeLa cells and a mammalian cell extract. We conclude that the nuclear import of DNA observed in these different vertebrate systems is largely mediated by the receptor transportin. We further report that histones, a known cargo of transportin, can act as an adaptor for the binding of transportin to DNA.
Delivery of DNA to the cell nucleus is an essential step in many types of viral infection, transfection, gene transfer by the plant pathogen Agrobacteriumtumefaciens and in strategies for gene therapy. Thus, the mechanism by which DNA crosses the nuclear pore complex (NPC) is of great interest. Using nuclei reconstituted in vitro inXenopus egg extracts, we previously studied DNA passage through the nuclear pores using a single-molecule approach based on optical tweezers. Fluorescently labeled DNA molecules were also seen to accumulate within nuclei. Here we find that this import of DNA relies on a soluble protein receptor of the importin family. To identify this receptor, we used different pathway-specific cargoes in competition studies as well as pathway-specific dominant negative inhibitors derived from the nucleoporin Nup153. We found that inhibition of the receptor transportin suppresses DNA import. In contrast, inhibition of importin β has little effect on the nuclear accumulation of DNA. The dependence on transportin was fully confirmed in assays using permeabilized HeLa cells and a mammalian cell extract. We conclude that the nuclear import of DNA observed in these different vertebrate systems is largely mediated by the receptor transportin. We further report that histones, a known cargo of transportin, can act as an adaptor for the binding of transportin to DNA.
Axially chiral facial amphiphiles with a dihydronaphthopentaphene structure as molecular tweezers for SWNTs
Renaud Marquis, Krystyna Kulikiewicz, Sergei Lebedkin, Manfred M. Kappes, Charles Mioskowski, Stéphane Meunier, Alain Wagner
Syntheses of chiral 6,15-dihydronaphtho[2,3-c]pentaphene derivatives of opposite configurations are reported. Starting from anthracene, the strategy involves two key steps: a Diels-Alder reaction on a prochiral dianthraquinone, and an enantiomeric resolution using (-)-menthol. The final molecules exhibit very strong optical activity, as shown by their circular dichroism spectra, and are examples of chiral facial amphiphiles. Their adsorption at the surface of single-walled carbon nanotubes (SWNTs) has also been studied, and has been found to occur preferentially on 0.8-1.0 nm diameter nanotubes among the population of a high-pressure CO conversion (HiPco) SWNT sample (0.8-1.2 nm). The synthesised facial amphiphiles act as nano-tweezers for the diameter-selective solubilisation of SWNTs in water. The expected optical activities of the SWNT samples solubilised by each of the chiral amphiphiles have been studied by circular dichroism spectroscopy, but the results are not yet conclusive.
Syntheses of chiral 6,15-dihydronaphtho[2,3-c]pentaphene derivatives of opposite configurations are reported. Starting from anthracene, the strategy involves two key steps: a Diels-Alder reaction on a prochiral dianthraquinone, and an enantiomeric resolution using (-)-menthol. The final molecules exhibit very strong optical activity, as shown by their circular dichroism spectra, and are examples of chiral facial amphiphiles. Their adsorption at the surface of single-walled carbon nanotubes (SWNTs) has also been studied, and has been found to occur preferentially on 0.8-1.0 nm diameter nanotubes among the population of a high-pressure CO conversion (HiPco) SWNT sample (0.8-1.2 nm). The synthesised facial amphiphiles act as nano-tweezers for the diameter-selective solubilisation of SWNTs in water. The expected optical activities of the SWNT samples solubilised by each of the chiral amphiphiles have been studied by circular dichroism spectroscopy, but the results are not yet conclusive.
Structural and micromechanical characterization of type I collagen gels
Olga Latinovic, Lawrence A. Hough and H. Daniel Ou-Yang
In this paper we report a study where we use a novel optical tweezers technique to measure the local viscoelastic properties of type I collagen solutions spanning the sol-to-gel transition. We use phase contrast optical microscopy to reveal dense and sparse regions of the rigid fibril networks, and find that the spatial variations in the mechanical properties of the collagen gels closely follow the structural properties. Within the dense phase of the connected network in the gel samples, there are regions that exhibit drastically different viscoelastic properties. Within the sparse regions of the gel samples, no evidence of elasticity is found. In type I collagen gels, we find a high degree of structural inhomogeneity. The inhomogeneity in the structural properties of collagen gels and the corresponding viscoelastic properties provide benchmark measurements for the behavior of desirable biological materials, or tissue equivalents.
In this paper we report a study where we use a novel optical tweezers technique to measure the local viscoelastic properties of type I collagen solutions spanning the sol-to-gel transition. We use phase contrast optical microscopy to reveal dense and sparse regions of the rigid fibril networks, and find that the spatial variations in the mechanical properties of the collagen gels closely follow the structural properties. Within the dense phase of the connected network in the gel samples, there are regions that exhibit drastically different viscoelastic properties. Within the sparse regions of the gel samples, no evidence of elasticity is found. In type I collagen gels, we find a high degree of structural inhomogeneity. The inhomogeneity in the structural properties of collagen gels and the corresponding viscoelastic properties provide benchmark measurements for the behavior of desirable biological materials, or tissue equivalents.
Unfolding and refolding properties of S pili on extraintestinal pathogenic Escherichia coli
Mickaël Castelain, Annika E. Sjöström, Erik Fällman, Bernt Eric Uhlin and Magnus Andersson
S pili are members of the chaperone-usher-pathway-assembled pili family that are predominantly associated with neonatal meningitis (SII) and believed to play a role in ascending urinary tract infections (SI). We used force-measuring optical tweezers to characterize the intrinsic biomechanical properties and kinetics of SII and SIpili. Under steady-state conditions, a sequential unfolding of the layers in the helix-like rod occurred at somewhat different forces, 26 pN for SII pili and 21 pN for SI pili, and there was an apparent difference in the kinetics, 1.3 and 8.8 Hz. Tests with bacteria defective in a newly recognized sfa gene (sfaX II) indicated that absence of the sfaX IIgene weakens the interactions of the fimbrium slightly and decreases the kinetics. Data of SI are compared with those of previously assessed pili primary associated with urinary tract infections, the P and type 1 pili. S pili have weaker layer-to-layer bonds than both P and type 1 pili, 21, 28 and 30 pN, respectively. In addition, the S pili kinetics are ~10 times faster than the kinetics of P pili and ~550 times faster than the kinetics of type 1 pili. Our results also show that the biomechanical properties of pili expressed ectopically from a plasmid in a laboratory strain (HB101) and pili expressed from the chromosome of a clinical isolate (IHE3034) are identical. Moreover, we demonstrate that it is possible to distinguish, by analyzing force-extension data, the different types of pili expressed by an individual cell of a clinical bacterial isolate.
S pili are members of the chaperone-usher-pathway-assembled pili family that are predominantly associated with neonatal meningitis (SII) and believed to play a role in ascending urinary tract infections (SI). We used force-measuring optical tweezers to characterize the intrinsic biomechanical properties and kinetics of SII and SIpili. Under steady-state conditions, a sequential unfolding of the layers in the helix-like rod occurred at somewhat different forces, 26 pN for SII pili and 21 pN for SI pili, and there was an apparent difference in the kinetics, 1.3 and 8.8 Hz. Tests with bacteria defective in a newly recognized sfa gene (sfaX II) indicated that absence of the sfaX IIgene weakens the interactions of the fimbrium slightly and decreases the kinetics. Data of SI are compared with those of previously assessed pili primary associated with urinary tract infections, the P and type 1 pili. S pili have weaker layer-to-layer bonds than both P and type 1 pili, 21, 28 and 30 pN, respectively. In addition, the S pili kinetics are ~10 times faster than the kinetics of P pili and ~550 times faster than the kinetics of type 1 pili. Our results also show that the biomechanical properties of pili expressed ectopically from a plasmid in a laboratory strain (HB101) and pili expressed from the chromosome of a clinical isolate (IHE3034) are identical. Moreover, we demonstrate that it is possible to distinguish, by analyzing force-extension data, the different types of pili expressed by an individual cell of a clinical bacterial isolate.
Impaired red cell deformability in iron deficient subjects
Marcelo M. Brandão, Maria de Lourdes R.B. Castro, Adriana Fontes, Carlos L. Cesar, Fernando F. Costa, Sara T.O. Saad
Iron deficiency is a systemic disorder, which affects a variety of different cell types and is one of the most frequent diseases throughout the world. The influence of iron deficiency upon erythrocyte deformability is controversial and could be a consequence of membrane peroxidation damage or cross linking of membrane proteins. The aim of this study was to determine the overall elasticity (the deformability of the entire cell is evaluated) of iron deficient red blood cells (RBC) using laser optical tweezers. In this study, the laser trapped the cell and the elasticity was then analyzed measuring cell deformation at six different drag velocities. Twenty-five RBCs from 11 healthy blood donors (controls) and 7 patients with iron deficiency anemia were analyzed. Iron deficiency subjects were classified into 3 groups based on Hb concentration for statistical analysis (group I: Hb = 7.0–7.9; group II: 8.0–10.2 and group III: 7.0–10.2 g/dl). The results showed an increased rigidity in the iron deficiency of deficient red blood cells when compared to normal control blood cells, and, this impaired deformability seems to be correlated to the hemoglobin concentration. In conclusion, the results obtained by optical tweezers showed that iron deficiency affects the elasticity of whole RBC.
Iron deficiency is a systemic disorder, which affects a variety of different cell types and is one of the most frequent diseases throughout the world. The influence of iron deficiency upon erythrocyte deformability is controversial and could be a consequence of membrane peroxidation damage or cross linking of membrane proteins. The aim of this study was to determine the overall elasticity (the deformability of the entire cell is evaluated) of iron deficient red blood cells (RBC) using laser optical tweezers. In this study, the laser trapped the cell and the elasticity was then analyzed measuring cell deformation at six different drag velocities. Twenty-five RBCs from 11 healthy blood donors (controls) and 7 patients with iron deficiency anemia were analyzed. Iron deficiency subjects were classified into 3 groups based on Hb concentration for statistical analysis (group I: Hb = 7.0–7.9; group II: 8.0–10.2 and group III: 7.0–10.2 g/dl). The results showed an increased rigidity in the iron deficiency of deficient red blood cells when compared to normal control blood cells, and, this impaired deformability seems to be correlated to the hemoglobin concentration. In conclusion, the results obtained by optical tweezers showed that iron deficiency affects the elasticity of whole RBC.
Hydrodynamic Mobility of an Optically Trapped Colloidal Particle near Fluid-Fluid Interfaces
G. M. Wang, R. Prabhakar, and E. M. Sevick
Using optical tweezers, we measure the anisotropic hydrodynamic mobility of a colloidal particle by tracking its thermal motion in an optical trap located near fluid-fluid interfaces, namely, liquid-vapor and liquid-liquid interfaces. The method requires no controlled fluid flow, is independent of conservative interactions between particle and interface, and resolves distance dependent friction to within a fraction of the particle radius. Near the liquid-vapor interface, the friction decreases below that in the bulk, corresponding to predictions of a “perfect-slip” surface.
Using optical tweezers, we measure the anisotropic hydrodynamic mobility of a colloidal particle by tracking its thermal motion in an optical trap located near fluid-fluid interfaces, namely, liquid-vapor and liquid-liquid interfaces. The method requires no controlled fluid flow, is independent of conservative interactions between particle and interface, and resolves distance dependent friction to within a fraction of the particle radius. Near the liquid-vapor interface, the friction decreases below that in the bulk, corresponding to predictions of a “perfect-slip” surface.
Combining Single-Molecule Optical Trapping and Small-Angle X-Ray Scattering Measurements to Compute the Persistence Length of a Protein ER/K alpha-Hel
S. Sivaramakrishnan, J. Sung, M. Ali, S. Doniach, H. Flyvbjerg and J.A. Spudich
A relatively unknown protein structure motif forms stable isolated single alpha-helices, termed ER/K alpha-helices, in a wide variety of proteins and has been shown to be essential for the function of some molecular motors. The flexibility of the ER/K alpha-helix determines whether it behaves as a force transducer, rigid spacer, or flexible linker in proteins. In this study, we quantity this flexibility in terms of persistence length, namely the length scale over which it is rigid. We use single-molecule optical trapping and small-angle x-ray scattering, combined with Monte Carlo simulations to demonstrate that the Kelch ER/K alpha-helix behaves as a wormlike chain with a persistence length of similar to 15 nm or similar to 28 turns of alpha-helix. The ER/K alpha-helix length in proteins varies from 3 to 60 nm, with a median length of similar to 5 nm. Knowledge of its persistence length enables us to define its function as a rigid spacer in a translation initiation factor, as a force transducer in the mechanoenzyme myosin VI, and as a flexible spacer in the Kelch-motif-containing protein.
Multi-particle three-dimensional coordinate estimation in real-time optical manipulation
J. S. Dam, I. Perch-Nielsen, D. Palima, J. Glückstad
We have previously shown how stereoscopic images can be obtained in our three-dimensional optical micromanipulation system [J. S. Dam et al, Opt. Express 16, 7244 (2008)]. Here, we present an extension and application of this principle to automatically gather the three-dimensional coordinates for all trapped particles with high tracking range and high reliability without requiring user calibration. Through deconvolving of the red, green, and blue colour planes to correct for bleeding between colour planes, we show that we can extend the system to also utilize green illumination, in addition to the blue and red. Applying the green colour as on-axis illumination yields redundant information for enhanced error correction, which is used to verify the gathered data, resulting in reliable coordinates as well as producing visually attractive images.
We have previously shown how stereoscopic images can be obtained in our three-dimensional optical micromanipulation system [J. S. Dam et al, Opt. Express 16, 7244 (2008)]. Here, we present an extension and application of this principle to automatically gather the three-dimensional coordinates for all trapped particles with high tracking range and high reliability without requiring user calibration. Through deconvolving of the red, green, and blue colour planes to correct for bleeding between colour planes, we show that we can extend the system to also utilize green illumination, in addition to the blue and red. Applying the green colour as on-axis illumination yields redundant information for enhanced error correction, which is used to verify the gathered data, resulting in reliable coordinates as well as producing visually attractive images.
Radiation force of scalar and electromagnetic twisted Gaussian Schell-model beams
Chengliang Zhao, Yangjian Cai, and Olga Korotkova
Radiation force of a focused scalar twisted Gaussian Schell-model (TGSM) beam on a Rayleigh dielectric sphere is investigated. It is found that the twist phase affects the radiation force and by raising the absolute value of the twist factor it is possible to increase both transverse and longitudinal trapping ranges at the real focus where the maximum on-axis intensity is located. Numerical calculations of radiation forces induced by a focused electromagnetic TGSM beam on a Rayleigh dielectric sphere are carried out. It is found that radiation force is closely related to the twist phase, degree of polarization and correlation factors of the initial beam. The trapping stability is also discussed.
Radiation force of a focused scalar twisted Gaussian Schell-model (TGSM) beam on a Rayleigh dielectric sphere is investigated. It is found that the twist phase affects the radiation force and by raising the absolute value of the twist factor it is possible to increase both transverse and longitudinal trapping ranges at the real focus where the maximum on-axis intensity is located. Numerical calculations of radiation forces induced by a focused electromagnetic TGSM beam on a Rayleigh dielectric sphere are carried out. It is found that radiation force is closely related to the twist phase, degree of polarization and correlation factors of the initial beam. The trapping stability is also discussed.
Monday, December 7, 2009
Optical forces on a Mie spheroidal particle arbitrarily oriented in a counterpropagating trap
Héctor Sosa-Martínez and Julio C. Gutiérrez-Vega
The axial and transverse optical forces exerted by Gaussian beams on an arbitrarily oriented and homogeneous spheroid are calculated and studied within the framework of the Mie theory. The results are applied to study the behavior of the forces in a counterpropagating optical trap. We calculate the trapping efficiencies for a wide range of physical parameters, including the beam waist separation distance, the equivalent spheroid radius, the spheroid eccentricity, and the refractive index ratio between the particle and the surrounding medium.
The axial and transverse optical forces exerted by Gaussian beams on an arbitrarily oriented and homogeneous spheroid are calculated and studied within the framework of the Mie theory. The results are applied to study the behavior of the forces in a counterpropagating optical trap. We calculate the trapping efficiencies for a wide range of physical parameters, including the beam waist separation distance, the equivalent spheroid radius, the spheroid eccentricity, and the refractive index ratio between the particle and the surrounding medium.
Manipulation and characterisation of accumulation and coarse mode aerosol particles using a Bessel beam trap
Helena Meresman, Jon B. Wills, Michael Summers, David McGloin and Jonathan P. Reid
Micron and sub-micron sized aerosol particles are captured, manipulated and characterised in a Bessel beam optical trap. Bright field microscopy and elastic light scattering measurements are used in combination to interrogate trapped particles and explore the optical landscape of the trap. We conclude that the Bessel trap has a number of advantages over optical tweezers in terms of characterization of accumulation mode particles, manipulation of particles over macroscopic length scales and effective control of the gas phase. As such, the Bessel trap is a valuable addition to the aerosol optical toolkit.
Micron and sub-micron sized aerosol particles are captured, manipulated and characterised in a Bessel beam optical trap. Bright field microscopy and elastic light scattering measurements are used in combination to interrogate trapped particles and explore the optical landscape of the trap. We conclude that the Bessel trap has a number of advantages over optical tweezers in terms of characterization of accumulation mode particles, manipulation of particles over macroscopic length scales and effective control of the gas phase. As such, the Bessel trap is a valuable addition to the aerosol optical toolkit.
Direct observation of the transfer of orbital angular momentum to metal particles from a focused circularly polarized Gaussian beam
Yiqiong Zhao, David Shapiro, David Mcgloin, Daniel T. Chiu, and Stefano Marchesini
It is well known that a circularly polarized Gaussian beam carries spin angular momentum, but not orbital angular momentum. This paper demonstrates that focusing a beam carrying spin angular momentum can induce an orbital angular momentum which we used to drive the orbital motion of a micron-sized metal particle that is trapped off the beam axis. The direction of the orbital motion is controlled by the handedness of the circular polarization. The orbiting dynamics of the trapped particle, which acted as an optical micro-detector, were quantitatively measured and found to be in excellent agreement with the theoretical predictions.
It is well known that a circularly polarized Gaussian beam carries spin angular momentum, but not orbital angular momentum. This paper demonstrates that focusing a beam carrying spin angular momentum can induce an orbital angular momentum which we used to drive the orbital motion of a micron-sized metal particle that is trapped off the beam axis. The direction of the orbital motion is controlled by the handedness of the circular polarization. The orbiting dynamics of the trapped particle, which acted as an optical micro-detector, were quantitatively measured and found to be in excellent agreement with the theoretical predictions.
Friday, December 4, 2009
Determining the Zero-Force Binding Energetics of an Intercalated DNA Complex by a Single-Molecule Approach
Tzu-Sen Yang, Yujia Cui, Chien-Ming Wu, Jem-Mau Lo, Chi-Shiun Chiang, Wun-Yi Shu, Wei-Ju Chung, Chung-Shan Yu, Kuo-Ning Chiang, Ian C. Hsu
Binding behavior of DNA: A wormlike chain model is applied to determine the zero-force binding energetics of an intercalated DNA complex. A mono-intercalating agent is synthesized which can be very easily inserted into the DNA bases (see figure). This approach is particularly important in nuclear medicine and new radiation therapies.
Binding behavior of DNA: A wormlike chain model is applied to determine the zero-force binding energetics of an intercalated DNA complex. A mono-intercalating agent is synthesized which can be very easily inserted into the DNA bases (see figure). This approach is particularly important in nuclear medicine and new radiation therapies.
Thursday, December 3, 2009
Steady electrical and micro-rheological response functions for uncharged colloidal inclusions in polyelectrolyte hydrogels
Aliasghar Mohammadi and Reghan J. Hill
The electric-field-induced response of an uncharged colloidal sphere embedded in a quenched polyelectrolyte hydrogel is calculated from a model where the polymer network is treated as an elastic, porous skeleton saturated with an aqueous electrolyte. We present exact analytical solutions for the steady response to a uniform electric field, as well as the steady susceptibility, defined as the ratio of the particle displacement to the strength of an optical or magnetic force. Even though the particle is uncharged, it attains a finite electric-field-induced displacement owing to hydrodynamic coupling with electroosmotic flow. The steady susceptibility decreases with increasing charge and decreasing electrolyte concentration; in general, charge imparts a small correction to the classical theory for an uncharged linearly elastic continuum.
Tuesday, December 1, 2009
Work distribution for a particle moving in an optical trap and non-Markovian bath
Alok Samanta, K. Srinivasu and Swapan K. Ghosh
We propose a simple approach to derive an exact analytical expression of work distribution for a system consisting of a colloidal particle trapped in an optical harmonic potential well, which is being pulled at a constant velocity through a solution represented by a non-Markovian bath. The thermal environment is represented by a bath composed of an infinite set of harmonic oscillators, and a model Hamiltonian for the trapped colloidal particle is constructed by representing the interaction with the bath via linear dissipative mechanism. We have studied the effects of pulling time, pulling speed, and the adiabatic limit. It is also observed that only at long time the total work is completely converted into dissipative work.
We propose a simple approach to derive an exact analytical expression of work distribution for a system consisting of a colloidal particle trapped in an optical harmonic potential well, which is being pulled at a constant velocity through a solution represented by a non-Markovian bath. The thermal environment is represented by a bath composed of an infinite set of harmonic oscillators, and a model Hamiltonian for the trapped colloidal particle is constructed by representing the interaction with the bath via linear dissipative mechanism. We have studied the effects of pulling time, pulling speed, and the adiabatic limit. It is also observed that only at long time the total work is completely converted into dissipative work.
Monday, November 30, 2009
Mode-selective thermal radiation from a microparticle
Hitoshi Odashima, Maki Tachikawa, and Kei Takehiro
We experimentally demonstrate that thermal radiation from a micron-sized dielectric particle depends sensitively on its size and shape through the cavity quantum-electrodynamic effect. Our laser trapping technique levitated a high-temperature microsphere of Al2O3 and enabled emission spectroscopy of the single particle. As the particle becomes smaller, a blackbody like spectrum turns into a spectrum dominated by multiple peaks resonant with whispering gallery modes of the spherical resonator. The observed sharp frequency selectivity is applicable to spectral control of thermal radiation.
We experimentally demonstrate that thermal radiation from a micron-sized dielectric particle depends sensitively on its size and shape through the cavity quantum-electrodynamic effect. Our laser trapping technique levitated a high-temperature microsphere of Al2O3 and enabled emission spectroscopy of the single particle. As the particle becomes smaller, a blackbody like spectrum turns into a spectrum dominated by multiple peaks resonant with whispering gallery modes of the spherical resonator. The observed sharp frequency selectivity is applicable to spectral control of thermal radiation.
Spontaneous symmetry breaking and circulation by optically bound microparticle chains in Gaussian beam traps
J. M. Taylor and G. D. Love
It has been known for some time that simple “optically bound” chains of dielectric microparticles can form in a counter propagating Gaussian beam optical trap. Here we report experimental observations of more complex trapped states, which do not reflect the underlying symmetry of the optical beam trap they are confined in. We discuss both stationary off-axis trapping and dynamic motion. We confirm the results using a rigorous Mie scattering model and also give a physical explanation for these static and dynamic off-axis trapped states.
It has been known for some time that simple “optically bound” chains of dielectric microparticles can form in a counter propagating Gaussian beam optical trap. Here we report experimental observations of more complex trapped states, which do not reflect the underlying symmetry of the optical beam trap they are confined in. We discuss both stationary off-axis trapping and dynamic motion. We confirm the results using a rigorous Mie scattering model and also give a physical explanation for these static and dynamic off-axis trapped states.
Design and realization of a high-stability optical tweezer
Slawomir Drobczynski, Pascal Hébraud, Jean Pierre Munch, and Sébastien Harlepp
We present a method in which we stabilize mechanically an optical tweezer setup over a period ranging up to 30 min. A feedback loop is used to correct the mechanical and thermal drifts. The position of a fixed object on the sample surface is measured with a CCD device and its fluctuations analyzed and used to maintain its position fixed with threepiezoelectric devices. With this setup, we obtain a spatial stability of 1.5 nm in the radial direction and 5 nm in the axial direction. This method opens the route for real-time measurements of kinetics of macromolecules association, at a single molecule level, on very long time scales.
We present a method in which we stabilize mechanically an optical tweezer setup over a period ranging up to 30 min. A feedback loop is used to correct the mechanical and thermal drifts. The position of a fixed object on the sample surface is measured with a CCD device and its fluctuations analyzed and used to maintain its position fixed with threepiezoelectric devices. With this setup, we obtain a spatial stability of 1.5 nm in the radial direction and 5 nm in the axial direction. This method opens the route for real-time measurements of kinetics of macromolecules association, at a single molecule level, on very long time scales.
Thursday, November 26, 2009
Binding Kinetics of Bisintercalator Triostin A with Optical Tweezers Force Mechanics
Christoph Kleimann, Andy Sischka, Andre Spiering, Katja Tönsing, Norbert Sewald, Ulf Diederichsen and Dario Anselmetti
The binding kinetics of the intercalative binding of Triostin A to λ-DNA was investigated by measuring the force extension response of the DNA-ligand complexes with an optical tweezers system. These force response curves, containing the information about different binding properties, were analyzed based on a recent method (put forth by another research group) for monointercalators that was extended to bisintercalators. Our binding analysis reveals an exponential dependence of the association constant on the applied external force as well as a decreasing binding site size. In general, our results are in agreement with those for the monointercalator ethidium. However, to explain the high-force binding site size, a new model for bisintercalation of Triostin A at high forces is proposed.
The binding kinetics of the intercalative binding of Triostin A to λ-DNA was investigated by measuring the force extension response of the DNA-ligand complexes with an optical tweezers system. These force response curves, containing the information about different binding properties, were analyzed based on a recent method (put forth by another research group) for monointercalators that was extended to bisintercalators. Our binding analysis reveals an exponential dependence of the association constant on the applied external force as well as a decreasing binding site size. In general, our results are in agreement with those for the monointercalator ethidium. However, to explain the high-force binding site size, a new model for bisintercalation of Triostin A at high forces is proposed.
Tug-of-war between dissimilar teams of microtubule motors regulates transport and fission of endosomes
Virupakshi Soppina, Arpan Kumar Rai, Avin Jayesh Ramaiya,Pradeep Barak and Roop Mallik
Intracellular transport is interspersed with frequent reversals in direction due to the presence of opposing kinesin and dynein motors on organelles that are carried as cargo. The cause and the mechanism of reversals are unknown, but are a key to understanding how cargos are delivered in a regulated manner to specific cellular locations. Unlike established single-motor biophysical assays, this problem requires understanding of the cooperative behavior of multiple interacting motors. Here we present measurements inside live Dictyosteliumcells, in a cell extract and with purified motors to quantify such an ensemble function of motors. We show through precise motion analysis that reversals during endosome motion are caused by a tug-of-war between kinesin and dynein. Further, we use a combination of optical trap-based force measurements and Monte Carlo simulations to make the surprising discovery that endosome transport uses many (approximately four to eight) weak and detachment-prone dyneins in a tug-of-war against a single strong and tenacious kinesin. We elucidate how this clever choice of dissimilar motors and motor teams achieves net transport together with endosome fission, both of which are important in controlling the balance of endocytic sorting. To the best of our knowledge, this is a unique demonstration that dynein and kinesin function differently at the molecular level inside cells and of how this difference is used in a specific cellular process, namely endosome biogenesis. Our work may provide a platform to understand intracellular transport of a variety of organelles in terms of measurable quantities.
Increasing trap stiffness with position clamping in holographic optical tweezers
Daryl Preece, Richard Bowman, Anna Linnenberger, Graham Gibson, Steven Serati, and Miles Padgett
We present a holographic optical tweezers system capable of position clamping multiple particles. Moving an optical trap in response to the trapped object’s motion is a powerful technique for optical control and force measurement. We have now realised this experimentally using a Boulder Nonlinear Systems Spatial Light Modulator (SLM) with a refresh rate of 203Hz. We obtain a reduction of 44% in the variance of the bead’s position, corresponding to an increase in effective trap stiffness of 77%. This reduction relies on the generation of holograms at high speed. We present software capable of calculating holograms in under 1ms using a graphics processor unit.
We present a holographic optical tweezers system capable of position clamping multiple particles. Moving an optical trap in response to the trapped object’s motion is a powerful technique for optical control and force measurement. We have now realised this experimentally using a Boulder Nonlinear Systems Spatial Light Modulator (SLM) with a refresh rate of 203Hz. We obtain a reduction of 44% in the variance of the bead’s position, corresponding to an increase in effective trap stiffness of 77%. This reduction relies on the generation of holograms at high speed. We present software capable of calculating holograms in under 1ms using a graphics processor unit.
Micro Manipulation with Optical Responsive Cholesteric and Compensated Nematic Liquid Crystal
Abu Kausar; Hiroto Nagano; Soichiro Okada; Tomonari Ogata; Seiji Kurihara
We have developed photoresponsive liquid crystalline films (cholesteric liquid crystal and compensated nematic liquid crystal) having azobenzene compound to manipulate micro particles on the surface by light irradiation. When few solid particles (polystyrene bead) were placed on the surface of the film, movement of the solid particles was observed upon irradiation of UV (365 nm) or visible light (>420 nm). Only rotational motion of the particles was observed on the cholesteric liquid crystal film. Both translation and rotation was observed on the compensated nematic liquid crystal film. So the mode of motion (either rotation or both translation and rotation) was changed by changing the film from cholesteric film to compensated liquid crystal film. The rotational direction of the solid was controlled by using chiral azobenzene compound with right or left handed twisting ability.
We have developed photoresponsive liquid crystalline films (cholesteric liquid crystal and compensated nematic liquid crystal) having azobenzene compound to manipulate micro particles on the surface by light irradiation. When few solid particles (polystyrene bead) were placed on the surface of the film, movement of the solid particles was observed upon irradiation of UV (365 nm) or visible light (>420 nm). Only rotational motion of the particles was observed on the cholesteric liquid crystal film. Both translation and rotation was observed on the compensated nematic liquid crystal film. So the mode of motion (either rotation or both translation and rotation) was changed by changing the film from cholesteric film to compensated liquid crystal film. The rotational direction of the solid was controlled by using chiral azobenzene compound with right or left handed twisting ability.
Tuesday, November 24, 2009
Forces between Blank Surfaces As Measured by the Colloidal Probe Technique and by Optical Tweezers − A Comparison
Mahdy M. Elmahdy, Astrid Drechsler, Christof Gutsche, Alla Synytska, Petra Uhlmann, Friedrich Kremer and Manfred Stamm
The well-established atomic force microscopy (AFM)-based colloidal probe technique (CPT) and optical tweezers (OT) are combined to measure the interaction forces between blank SiO2 surfaces in aqueous ionic solutions (CaCl2) of varying concentration at pH 7. Spherical colloids (SiO2, diameter 4.63 ± 0.05 μm) taken out of the same batch are used by both methods. In the case of CPT, a single colloid is glued to a cantilever, and the interaction forces with a plain SiO2 surface are determined in dependence on the concentration of the surrounding medium. For the OT studies, two colloids (one fixed to a micropipet by capillary action, the other held with the optical trap) are approached to each other in nanometer steps, and the resulting forces are measured for the same media as in the CPT experiment. Both techniques fit well to each other and enable one to cover interaction energies ranging from 10−5 to 1 mN/m. The experimental data are well described by the Derjaguin−Landau−Verwey−Overbeek (DLVO) theory revealing that the effective surface charge density changes slightly with concentration.
The well-established atomic force microscopy (AFM)-based colloidal probe technique (CPT) and optical tweezers (OT) are combined to measure the interaction forces between blank SiO2 surfaces in aqueous ionic solutions (CaCl2) of varying concentration at pH 7. Spherical colloids (SiO2, diameter 4.63 ± 0.05 μm) taken out of the same batch are used by both methods. In the case of CPT, a single colloid is glued to a cantilever, and the interaction forces with a plain SiO2 surface are determined in dependence on the concentration of the surrounding medium. For the OT studies, two colloids (one fixed to a micropipet by capillary action, the other held with the optical trap) are approached to each other in nanometer steps, and the resulting forces are measured for the same media as in the CPT experiment. Both techniques fit well to each other and enable one to cover interaction energies ranging from 10−5 to 1 mN/m. The experimental data are well described by the Derjaguin−Landau−Verwey−Overbeek (DLVO) theory revealing that the effective surface charge density changes slightly with concentration.
Determination of cell elasticity through hybrid ray optics and continuum mechanics modeling of cell deformation in the optical stretcher
Andrew E. Ekpenyong, Carolyn L. Posey, Joy L. Chaput, Anya K. Burkart, Meg M. Marquardt, Timothy J. Smith, and Michael G. Nichols
The optical stretcher is a dual-beam trap capable of stretching individual cells. Previous studies have used either ray-or wave-optical models to compute the optical pressure on the surface of a spherical cell. We have extended the ray-optics model to account for focusing by the spherical interface and the effects of multiple internal reflections. Simulation results for red-blood cells (RBCs) show that internal reflections can lead to significant perturbation of the deformation, leading to a systematic error in the determination of cellular elasticity. Calibration studies show excellent agreement between the predicted and measured escape force, and RBC stiffness measurements are consistent with literature values. Measurements of the elasticity of murine osteogenic cells reveal that these cells are approximately 5.4 times stiffer than RBCs.
The optical stretcher is a dual-beam trap capable of stretching individual cells. Previous studies have used either ray-or wave-optical models to compute the optical pressure on the surface of a spherical cell. We have extended the ray-optics model to account for focusing by the spherical interface and the effects of multiple internal reflections. Simulation results for red-blood cells (RBCs) show that internal reflections can lead to significant perturbation of the deformation, leading to a systematic error in the determination of cellular elasticity. Calibration studies show excellent agreement between the predicted and measured escape force, and RBC stiffness measurements are consistent with literature values. Measurements of the elasticity of murine osteogenic cells reveal that these cells are approximately 5.4 times stiffer than RBCs.
Microfluidic sorting system based on optical force switching
S.-K. Hoi, C. Udalagama, C.-H. Sow, F. Watt and A. A. Bettiol
We report a versatile, and automatic method for sorting cells and particles in a three dimensional polydimethylsiloxane (PDMS) structure consisting of two cross-microchannels. As microspheres or yeast cells are fed continuously into a lower channel, a line shaped focused laser beam is applied (perpendicular to the direction of flow) at the crossing junction of the two channels. The scattering force of the laser beam was employed to push microparticles matching specific criteria upwards from one channel to another. The force depends on the intrinsic properties of the particles such as their refractive index and size, as well as the laser power and the fluid flow speed. The combination of these parameters gives a tunable selection criterion for the effective and efficient sorting of the particles. The introduction of the cylindrical lens into the optical train allows for simultaneous manipulation of multiple particles which has significantly increased the efficiency and throughput of the sorting. A high aspect ratio microchannel (A.R.=1.6) was found to enhance the sorting performance of the device. By careful control of the microparticle flow rate, near 100% sorting efficiency was achieved.
We report a versatile, and automatic method for sorting cells and particles in a three dimensional polydimethylsiloxane (PDMS) structure consisting of two cross-microchannels. As microspheres or yeast cells are fed continuously into a lower channel, a line shaped focused laser beam is applied (perpendicular to the direction of flow) at the crossing junction of the two channels. The scattering force of the laser beam was employed to push microparticles matching specific criteria upwards from one channel to another. The force depends on the intrinsic properties of the particles such as their refractive index and size, as well as the laser power and the fluid flow speed. The combination of these parameters gives a tunable selection criterion for the effective and efficient sorting of the particles. The introduction of the cylindrical lens into the optical train allows for simultaneous manipulation of multiple particles which has significantly increased the efficiency and throughput of the sorting. A high aspect ratio microchannel (A.R.=1.6) was found to enhance the sorting performance of the device. By careful control of the microparticle flow rate, near 100% sorting efficiency was achieved.
Forces of Interaction between Poly(2-vinylpyridine) Brushes As Measured by Optical Tweezers
Mahdy M. Elmahdy, Alla Synytska, Astrid Drechsler, Christof Gutsche, Petra Uhlmann, Manfred Stamm and Friedrich Kremer
Forces of interaction within single pairs of poly(2-vinylpyridine) (P2VP) grafted colloids have been measured by optical tweezers (OT) with an extraordinary resolution of ±0.5 pN. Parameters to be varied are the concentration and type of salt (KCl, CaCl2, and LaCl3) of the surrounding medium as well as its pH. The observed force−distance relation is quantitativelydescribed by the Jusufi model [Colloid Polym. Sci. 2004, 282, 910−917] for spherical polyelectrolyte brushes which takes into account the entropic effect of the counterions and enables one to estimate the ionic concentration inside the brush. The transition from an osmotic to the salted brush regime is analyzed in detail. For the scaling of the brush height a power law is found having an exponent of 0.24 ± 0.01, which ranges between the values expected for spherical and planar brushes. At pH 4 a strong transition from a brush to a pancake conformation takes place.
Light-Induced Nonlinear Rotations of Nematic Liquid Crystal Droplets Trapped in Laser Tweezers
Etienne Brasselet, Tadas Balciunas; Naoki Murazawa; Saulius Juodkazis; Hiroaki Misawa
We report on optically induced rotations of nematic liquid crystal droplets. We show experimentally that bipolar nematic liquid crystal droplets trapped in laser tweezers having circular polarization can exhibit nonlinear rotational motion, unlike optically trapped solid birefringent micro-plates. Such nonlinear rotations are retrieved by analyzing the polarization dynamics analysis of the light beam after it has passed through the droplet. The occurrence of complex dynamics is only found for large enough trapping power and droplet diameter. The analogy with optically induced nonlinear rotations in nematic liquid crystal films is briefly discussed.
We report on optically induced rotations of nematic liquid crystal droplets. We show experimentally that bipolar nematic liquid crystal droplets trapped in laser tweezers having circular polarization can exhibit nonlinear rotational motion, unlike optically trapped solid birefringent micro-plates. Such nonlinear rotations are retrieved by analyzing the polarization dynamics analysis of the light beam after it has passed through the droplet. The occurrence of complex dynamics is only found for large enough trapping power and droplet diameter. The analogy with optically induced nonlinear rotations in nematic liquid crystal films is briefly discussed.
Thursday, November 19, 2009
Precise balancing of viscous and radiation forces on a particle in liquid-filled photonic bandgap fiber
T. G. Euser, M. K. Garbos, J. S. Y. Chen, and P. St.J. Russell
A great challenge in microfluidics is the precise control of laser radiation forces acting on single particles or cells, while allowing monitoring of their optical and chemical properties. We show that, in the liquid-filled hollow core of a single-mode photonic crystal fiber, a micrometer-sized particle can be held stably against a fluidic counterflow using radiation pressure and can be moved to and fro (over tens of centimeters) by ramping the laser power up and down. Accurate studies of the microfluidic drag forces become possible, because the particle is trapped in the center of the single guided optical mode, resulting in highly reproducible radiation forces. The counterflowing liquid can be loaded with sequences of chemicals in precisely controlled concentrations and doses, making possible studies of single particles, vesicles, or cells.
A great challenge in microfluidics is the precise control of laser radiation forces acting on single particles or cells, while allowing monitoring of their optical and chemical properties. We show that, in the liquid-filled hollow core of a single-mode photonic crystal fiber, a micrometer-sized particle can be held stably against a fluidic counterflow using radiation pressure and can be moved to and fro (over tens of centimeters) by ramping the laser power up and down. Accurate studies of the microfluidic drag forces become possible, because the particle is trapped in the center of the single guided optical mode, resulting in highly reproducible radiation forces. The counterflowing liquid can be loaded with sequences of chemicals in precisely controlled concentrations and doses, making possible studies of single particles, vesicles, or cells.
Tuesday, November 17, 2009
Trapping double negative particles in the ray optics regime using optical tweezers with focused beams
Leonardo A. Ambrosio and H. E. Hernández-Figueroa
The capabilities of optical tweezers to trap DNG (double negative) spherical particles, with both negative permittivity and permeability, are explored in detail by analyzing some interesting theoretical features not seeing in conventional DPS (double positive) particles possessing positive refractive index. The ray optics regime is adopted and, although this regime is quite simple and limited, its validity is already known and tested for DPS particles such as biological cells and molecules trapped by highly focused beams. Simulation results confirm that even for ray optics, DNG particles present unusual and interesting trapping characteristics.
The capabilities of optical tweezers to trap DNG (double negative) spherical particles, with both negative permittivity and permeability, are explored in detail by analyzing some interesting theoretical features not seeing in conventional DPS (double positive) particles possessing positive refractive index. The ray optics regime is adopted and, although this regime is quite simple and limited, its validity is already known and tested for DPS particles such as biological cells and molecules trapped by highly focused beams. Simulation results confirm that even for ray optics, DNG particles present unusual and interesting trapping characteristics.
Highly birefringent vaterite microspheres: production, characterization and applications for optical micromanipulation
Simon J. Parkin, Robert Vogel, Martin Persson, Maren Funk, Vincent L. Loke, Timo A. Nieminen, Norman R. Heckenberg, and Halina Rubinsztein-Dunlop
This paper reports on a simple synthesis and characterization of highly birefringent vaterite microspheres, which are composed of 20–30 nm sized nanocrystalls. Scanning electron microscopy shows a quite disordered assembly of nanocrystals within the microspheres. However, using optical tweezers, the effective birefringence of the microspheres was measured to be Δn = 0.06, which compares to Δn = 0.1 of vaterite single crystals. This suggests a very high orientation of the nanocrystals within the microspheres. A hyperbolic model of the direction of the optical axis throughout the vaterite spherulite best fits the experimental data. Results from polarized light microscopy further confirm the hyperbolic model.
This paper reports on a simple synthesis and characterization of highly birefringent vaterite microspheres, which are composed of 20–30 nm sized nanocrystalls. Scanning electron microscopy shows a quite disordered assembly of nanocrystals within the microspheres. However, using optical tweezers, the effective birefringence of the microspheres was measured to be Δn = 0.06, which compares to Δn = 0.1 of vaterite single crystals. This suggests a very high orientation of the nanocrystals within the microspheres. A hyperbolic model of the direction of the optical axis throughout the vaterite spherulite best fits the experimental data. Results from polarized light microscopy further confirm the hyperbolic model.
Monday, November 16, 2009
Forces in nematic liquid crystals: from nanoscale interfacial forces to long-range forces in nematic colloids
Igor Mu
evi
evi In this paper we give an overview of experiments that provided an insight into the nature of forces between surfaces and objects in a nematic liquid crystal. These forces, also called 'structural forces', are the consequence of the long-range orientational order and orientational elasticity of nematic liquid crystals. Owing to their fundamental as well as technological importance, forces between objects in liquid crystals have been a subject of growing interest during the last decade. Experimental observations and studies of structural forces are described from nanoscale interfacial forces, measured by an atomic force microscope, to the micro-scale forces between colloidal particles in nematics, studied by laser tweezers and optical video microscopy.
An optical-manipulation technique for cells in physiological flows
Hu Zhang, Neng H. Chen, Alicia El Haj and Kuo-Kang Liu
We have developed a technique to manipulate human red blood cells (RBCs) in hydrodynamic flows. This method applies optical tweezers to trap and move microbead-attached RBCs in a liquid medium at various speeds, while it significantly minimizes laser heating and photon-induced stress for normal operation with laser-trapped cells. Computational fluid dynamics is applied to simulate flow-induced shear stress over the cell membrane and to correlate quantitatively the forces with the cell deformations. RBCs can be manipulated under physiological conditions by this approach, which may open an avenue to design principles for the next generation of cell sorting and delivery.
We have developed a technique to manipulate human red blood cells (RBCs) in hydrodynamic flows. This method applies optical tweezers to trap and move microbead-attached RBCs in a liquid medium at various speeds, while it significantly minimizes laser heating and photon-induced stress for normal operation with laser-trapped cells. Computational fluid dynamics is applied to simulate flow-induced shear stress over the cell membrane and to correlate quantitatively the forces with the cell deformations. RBCs can be manipulated under physiological conditions by this approach, which may open an avenue to design principles for the next generation of cell sorting and delivery.
High-resolution, long-term characterization of bacterial motility using optical tweezers
Taejin L Min, Patrick J Mears, Lon M Chubiz, Christopher V Rao, Ido Golding & Yann R Chemla
We present a single-cell motility assay, which allows the quantification of bacterial swimming in a well-controlled environment, for durations of up to an hour and with a temporal resolution greater than the flagellar rotation rates of 100 Hz. The assay is based on an instrument combining optical tweezers, light and fluorescence microscopy, and a microfluidic chamber. Using this device we characterized the long-term statistics of the run-tumble time series in individual Escherichia coli cells. We also quantified higher-order features of bacterial swimming, such as changes in velocity and reversals of swimming direction.
We present a single-cell motility assay, which allows the quantification of bacterial swimming in a well-controlled environment, for durations of up to an hour and with a temporal resolution greater than the flagellar rotation rates of 100 Hz. The assay is based on an instrument combining optical tweezers, light and fluorescence microscopy, and a microfluidic chamber. Using this device we characterized the long-term statistics of the run-tumble time series in individual Escherichia coli cells. We also quantified higher-order features of bacterial swimming, such as changes in velocity and reversals of swimming direction.
Parallel trapping of multiwalled carbon nanotubes with optoelectronic tweezers
Peter J. Pauzauskie, Arash Jamshidi, Justin K. Valley, Joe H. Satcher, Jr., and Ming C. Wu
Here we report the use of optoelectronic tweezers and dynamic virtual electrodes to address multiwalled carbon nanotubes (MWCNTs) with trap stiffness values of approximately 50 fN/µm. Both high-speed translation (>200 µm/s) of individual-MWCNTsand two-dimensional trapping of MWCNT ensembles are achieved using 100,000 times less optical power density than single beam laser tweezers. Modulating the virtual electrode's intensity enables tuning of the MWCNT ensemble's number density by an order of magnitude on the time scale of seconds promising a broad range of applications in MWCNT science and technology.
Here we report the use of optoelectronic tweezers and dynamic virtual electrodes to address multiwalled carbon nanotubes (MWCNTs) with trap stiffness values of approximately 50 fN/µm. Both high-speed translation (>200 µm/s) of individual-MWCNTsand two-dimensional trapping of MWCNT ensembles are achieved using 100,000 times less optical power density than single beam laser tweezers. Modulating the virtual electrode's intensity enables tuning of the MWCNT ensemble's number density by an order of magnitude on the time scale of seconds promising a broad range of applications in MWCNT science and technology.
Amplitude and frequency spectra of thermal fluctuations of a translocating RNA molecule
Henk Vocks, Debabrata Panja and Gerard T Barkema
Using a combination of theory and computer simulations, we study the translocation of an RNA molecule, pulled through a solid-state nanopore by an optical tweezer, as a method for determining its secondary structure. The resolution with which the elements of the secondary structure can be determined is limited by thermal fluctuations. We present a detailed study of these thermal fluctuations, including the frequency spectrum, and show that these rule out single-nucleotide resolution under the experimental conditions which we simulated. Two possible ways to improve this resolution are strongly stretching the RNA with a back-pulling voltage across the membrane, and stiffening the translocated part of the RNA by biochemical means.
Using a combination of theory and computer simulations, we study the translocation of an RNA molecule, pulled through a solid-state nanopore by an optical tweezer, as a method for determining its secondary structure. The resolution with which the elements of the secondary structure can be determined is limited by thermal fluctuations. We present a detailed study of these thermal fluctuations, including the frequency spectrum, and show that these rule out single-nucleotide resolution under the experimental conditions which we simulated. Two possible ways to improve this resolution are strongly stretching the RNA with a back-pulling voltage across the membrane, and stiffening the translocated part of the RNA by biochemical means.
Friday, November 13, 2009
Multiple traps created with an inclined dual-fiber system
Yuxiang Liu and Miao Yu
Multiple optical traps allow one to manipulate multiple particles simultaneously, to characterize interactions in colloidal systems, and to assemble particles into complex structures. Most of the current multiple optical traps are realized with microscope objective-based optical tweezers, which are bulky in size. In this article, we created multiple optical traps with an inclined dual-fiber optical tweezers setup. One 3D trap and two 2D traps were formed at different vertical levels with adjustable separations and positions. We demonstrated that this fiber-based trapping system can be used as a simple block to perform multiple functions, such as particle grouping, separation, and stacking. Moreover, we found that multiple beads can be trapped and stacked up in three dimensions. Compared with those formed with objective-based optical tweezers, the multiple traps presented here are small in size and independent of the objective or the substrate, and hence hold the promise to be integrated in microfluidic systems. This fiber-based multiple traps can be used for on-chip parallel manipulation, particle separation, and characterization of interactions of colloidal and biological systems.
Multiple optical traps allow one to manipulate multiple particles simultaneously, to characterize interactions in colloidal systems, and to assemble particles into complex structures. Most of the current multiple optical traps are realized with microscope objective-based optical tweezers, which are bulky in size. In this article, we created multiple optical traps with an inclined dual-fiber optical tweezers setup. One 3D trap and two 2D traps were formed at different vertical levels with adjustable separations and positions. We demonstrated that this fiber-based trapping system can be used as a simple block to perform multiple functions, such as particle grouping, separation, and stacking. Moreover, we found that multiple beads can be trapped and stacked up in three dimensions. Compared with those formed with objective-based optical tweezers, the multiple traps presented here are small in size and independent of the objective or the substrate, and hence hold the promise to be integrated in microfluidic systems. This fiber-based multiple traps can be used for on-chip parallel manipulation, particle separation, and characterization of interactions of colloidal and biological systems.
Singular optical manipulation of birefringent elastic media using nonsingular beams
Etienne Brasselet
It is shown that nonsingular light beams can generate singular birefringent patterns in homogeneous birefringent elastic media. These orientational defects of the optical-axis spatial distribution originate from an optical torque driven by a nonzero longitudinal field component. Singular radial and spin-dependent azimuthal light-induced elastic distortion patterns are described and experimentally observed in a uniform liquid-crystal film in the course of a focused circularly polarized Gaussian beam.
It is shown that nonsingular light beams can generate singular birefringent patterns in homogeneous birefringent elastic media. These orientational defects of the optical-axis spatial distribution originate from an optical torque driven by a nonzero longitudinal field component. Singular radial and spin-dependent azimuthal light-induced elastic distortion patterns are described and experimentally observed in a uniform liquid-crystal film in the course of a focused circularly polarized Gaussian beam.
Stable optical trapping of latex nanoparticles with ultrashort pulsed illumination
Arijit Kumar De, Debjit Roy, Aveek Dutta, and Debabrata Goswami
Here we report how ultrafast pulsed illumination at low average power results in a stable three-dimensional (3D) optical trap holding latex nanoparticles which is otherwise not possible with continuous wave lasers at the same power level. The gigantic peak power of a femtosecond pulse exerts a huge instantaneous gradient force that has been predicted theoretically earlier and implemented for microsecond pulses in a different context by others. In addition, the resulting two-photon fluorescence allows direct observation of trapping events by providing intrinsic 3D resolution.
Here we report how ultrafast pulsed illumination at low average power results in a stable three-dimensional (3D) optical trap holding latex nanoparticles which is otherwise not possible with continuous wave lasers at the same power level. The gigantic peak power of a femtosecond pulse exerts a huge instantaneous gradient force that has been predicted theoretically earlier and implemented for microsecond pulses in a different context by others. In addition, the resulting two-photon fluorescence allows direct observation of trapping events by providing intrinsic 3D resolution.
Experimental analysis of recoil effects induced by fluorescence photons
Alexander Zhdanov, Satish Rao, Andrey Fedyanin, and Dmitri Petrov
The momentum transfer to a scatterer from fluorescence photons was detected using an optical system that permits one to simultaneously measure the radiation force exerted on and fluorescence emission from the scatterer. The core of this technique is a partially metal covered dielectric bead optically trapped in a liquid with dye molecules. Fluorescence emission from the volume that includes the bead is measured simultaneously with the Brownian motion of the bead. The perturbed motion of the bead is a result of photon momentum transfer from the fluorescence of the dye to the trapped scatterer. The bead position fluctuations indicate the presence of the fluorescence and its bleaching nature. The results demonstrate the capability of the photonic force microscopy technique to be a complement to spectroscopy in the study of optical processes.
The momentum transfer to a scatterer from fluorescence photons was detected using an optical system that permits one to simultaneously measure the radiation force exerted on and fluorescence emission from the scatterer. The core of this technique is a partially metal covered dielectric bead optically trapped in a liquid with dye molecules. Fluorescence emission from the volume that includes the bead is measured simultaneously with the Brownian motion of the bead. The perturbed motion of the bead is a result of photon momentum transfer from the fluorescence of the dye to the trapped scatterer. The bead position fluctuations indicate the presence of the fluorescence and its bleaching nature. The results demonstrate the capability of the photonic force microscopy technique to be a complement to spectroscopy in the study of optical processes.
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