Ying Li, Yanjun Hu
This work demonstrates optical trapping and transportation of nanoparticles along a nanofiber microring. The optical fiber, 800 nm in diameter, was formed to be a microring with a bending radius of about 42 μm. A microparticle, stuck to the junction of the ring, plays a critical role for confining nanoparticles on the microring. The experimental results show that, when a 650 nm laser light was launched into the ring, nanoparticles dispersed in the solution were trapped to the ring surface and delivered along the direction of the light propagation. Because the stuck microparticle on the junction for perturbation, nanoparticles can be confined over entire microring circumference. This technology offers a new degree of control for particles and lead to various nanomanipulation applictions.
DOI
Tuesday, January 30, 2018
Gold nanocups fabricated using two-dimensional colloidal crystals and simulation of their optical trapping force
Hiroaki Agawa,Takayuki Okamoto, Toshihiro Isobe, Akira Nakajima, and Sachiko Matsushita
Optical tweezers are powerful and flexible tools for manipulating micrometre-sized objects. Recently, metal nanostructures are gathering attention to trap nanometre-sized objects to utilize their plasmonic properties. Here, we discuss the preparation of gold nanocups (250-nm diameter) and their plasmonic properties for optical trapping. Gold was sputtered on a two-dimensional colloidal crystal (2DCC) and then de-coupled from the 2DCC via hydrofluoric acid etching, which resulted in a uniform gold nanocup array that was encapsulated in a flexible polymer resin. These nanocups in a resin can be transferred onto a variety of substrates. The optical trapping properties of gold nanocups are also discussed.
DOI
Optical tweezers are powerful and flexible tools for manipulating micrometre-sized objects. Recently, metal nanostructures are gathering attention to trap nanometre-sized objects to utilize their plasmonic properties. Here, we discuss the preparation of gold nanocups (250-nm diameter) and their plasmonic properties for optical trapping. Gold was sputtered on a two-dimensional colloidal crystal (2DCC) and then de-coupled from the 2DCC via hydrofluoric acid etching, which resulted in a uniform gold nanocup array that was encapsulated in a flexible polymer resin. These nanocups in a resin can be transferred onto a variety of substrates. The optical trapping properties of gold nanocups are also discussed.
DOI
Calcium effect on membrane of an optically trapped erythrocyte studied by digital holographic microscopy
Vahideh Farzam Rad, Rahim Tavakkoli, Ali-Reza Moradi, Arun Anand, and Bahram Javidi
The calcium level in blood affects the morphological and rheological properties of red blood cell (RBC) membranes. In this paper, we present an integrated optical system for a single cell study of hypercalcemia. The system consists of holographic optical tweezers and blinking optical tweezers, for photo-damage-free immobilization of the cells, combined with digital holographic microscopy, for quantitative analysis and live visualization of the cells. Digital holograms were recorded live, while the concentration of calcium ions in the buffer is gradually increased. Full morphometric data of RBCs were obtained by numerical reconstruction of the holograms. Morphological changes are expressed in terms of various parameters such as root mean square, skewness, and kurtosis of the cell membrane thickness distribution. We have observed dramatic changes of the cell morphology, which are attributed to the formation of calcium-induced hydrophobic aggregates of phospholipid molecules in the RBC membrane, resulting in a net change in membrane rigidity. Our experimental results are in agreement with previous biological studies of RBCs under the Ca2+ influence.
DOI
The calcium level in blood affects the morphological and rheological properties of red blood cell (RBC) membranes. In this paper, we present an integrated optical system for a single cell study of hypercalcemia. The system consists of holographic optical tweezers and blinking optical tweezers, for photo-damage-free immobilization of the cells, combined with digital holographic microscopy, for quantitative analysis and live visualization of the cells. Digital holograms were recorded live, while the concentration of calcium ions in the buffer is gradually increased. Full morphometric data of RBCs were obtained by numerical reconstruction of the holograms. Morphological changes are expressed in terms of various parameters such as root mean square, skewness, and kurtosis of the cell membrane thickness distribution. We have observed dramatic changes of the cell morphology, which are attributed to the formation of calcium-induced hydrophobic aggregates of phospholipid molecules in the RBC membrane, resulting in a net change in membrane rigidity. Our experimental results are in agreement with previous biological studies of RBCs under the Ca2+ influence.
DOI
Depth-resolved measurement of optical radiation-pressure forces with optical coherence tomography
Nichaluk Leartprapun, Rishyashring R. Iyer, and Steven G. Adie
A weakly focused laser beam can exert sufficient radiation pressure to manipulate microscopic particles over a large depth range. However, depth-resolved continuous measurement of radiation-pressure force profiles over an extended range about the focal plane has not been demonstrated despite decades of research on optical manipulation. Here, we present a method for continuous measurement of axial radiation-pressure forces from a weakly focused beam on polystyrene micro-beads suspended in viscous fluids over a depth range of 400 μm, based on real-time monitoring of particle dynamics using optical coherence tomography (OCT). Measurements of radiation-pressure forces as a function of beam power, wavelength, bead size, and refractive index are consistent with theoretical trends. However, our continuous measurements also reveal localized depth-dependent features in the radiation-pressure force profiles that deviate from theoretical predictions based on an aberration-free Gaussian beam. The combination of long-range radiation pressure and OCT offers a new mode of quantitative optical manipulation and detection with extended spatial coverage. This may find applications in the characterization of optical tractor beams, or volumetric optical manipulation and interrogation of beads in viscoelastic media.
DOI
A weakly focused laser beam can exert sufficient radiation pressure to manipulate microscopic particles over a large depth range. However, depth-resolved continuous measurement of radiation-pressure force profiles over an extended range about the focal plane has not been demonstrated despite decades of research on optical manipulation. Here, we present a method for continuous measurement of axial radiation-pressure forces from a weakly focused beam on polystyrene micro-beads suspended in viscous fluids over a depth range of 400 μm, based on real-time monitoring of particle dynamics using optical coherence tomography (OCT). Measurements of radiation-pressure forces as a function of beam power, wavelength, bead size, and refractive index are consistent with theoretical trends. However, our continuous measurements also reveal localized depth-dependent features in the radiation-pressure force profiles that deviate from theoretical predictions based on an aberration-free Gaussian beam. The combination of long-range radiation pressure and OCT offers a new mode of quantitative optical manipulation and detection with extended spatial coverage. This may find applications in the characterization of optical tractor beams, or volumetric optical manipulation and interrogation of beads in viscoelastic media.
DOI
Microsystems for Single-Cell Analysis
Sonja M. Weiz, Mariana Medina-Sánchez, Oliver G. Schmidt
Due to the heterogeneity that exists even between cells of the same tissue, it is essential to use techniques and devices able to resolve the characteristics of single biological cells, such as morphology, metabolism, or response to drugs. To that end, different structures with sizes similar to that of individual cells have been developed in recent years, which allow single-cell studies with high sensitivity and high resolution. By employing a variety of sensing strategies, one can obtain complementary information about individual cells, and thus create a complete picture of cellular properties. This review aims to provide an overview of microscale single-cell sensors. The progress in micrometer-sized sensing probes as well as microfluidic and micropatterned devices is described, showing the capabilities of the available systems. In addition, a comprehensive compendium of systems based on rolled-up microtubes, which have the potential to advance and improve the single-cell analysis microsystem field, is comprised.
DOI
Due to the heterogeneity that exists even between cells of the same tissue, it is essential to use techniques and devices able to resolve the characteristics of single biological cells, such as morphology, metabolism, or response to drugs. To that end, different structures with sizes similar to that of individual cells have been developed in recent years, which allow single-cell studies with high sensitivity and high resolution. By employing a variety of sensing strategies, one can obtain complementary information about individual cells, and thus create a complete picture of cellular properties. This review aims to provide an overview of microscale single-cell sensors. The progress in micrometer-sized sensing probes as well as microfluidic and micropatterned devices is described, showing the capabilities of the available systems. In addition, a comprehensive compendium of systems based on rolled-up microtubes, which have the potential to advance and improve the single-cell analysis microsystem field, is comprised.
DOI
Nematic Liquid-Crystal Colloids
Igor Muševič
This article provides a concise review of a new state of colloidal matter called nematic liquid-crystal colloids. These colloids are obtained by dispersing microparticles of different shapes in a nematic liquid crystal that acts as a solvent for the dispersed particles. The microparticles induce a local deformation of the liquid crystal, which then generates topological defects and long-range forces between the neighboring particles. The colloidal forces in nematic colloids are much stronger than the forces in ordinary colloids in isotropic solvents, exceeding thousands of kBT per micrometer-sized particle. Of special interest are the topological defects in nematic colloids, which appear in many fascinating forms, such as singular points, closed loops, multitudes of interlinked and knotted loops or soliton-like structures. The richness of the topological phenomena and the possibility to design and control topological defects with laser tweezers make colloids in nematic liquid crystals an excellent playground for testing the basic theorems of topology.
DOI
This article provides a concise review of a new state of colloidal matter called nematic liquid-crystal colloids. These colloids are obtained by dispersing microparticles of different shapes in a nematic liquid crystal that acts as a solvent for the dispersed particles. The microparticles induce a local deformation of the liquid crystal, which then generates topological defects and long-range forces between the neighboring particles. The colloidal forces in nematic colloids are much stronger than the forces in ordinary colloids in isotropic solvents, exceeding thousands of kBT per micrometer-sized particle. Of special interest are the topological defects in nematic colloids, which appear in many fascinating forms, such as singular points, closed loops, multitudes of interlinked and knotted loops or soliton-like structures. The richness of the topological phenomena and the possibility to design and control topological defects with laser tweezers make colloids in nematic liquid crystals an excellent playground for testing the basic theorems of topology.
DOI
Thursday, January 25, 2018
Folding and Domain Interactions of Three Orthologs of Hsp90 Studied by Single-Molecule Force Spectroscopy
Markus Jahn, Katarzyna Tych, Hannah Girstmair, Maximilian Steinmaßl, Thorsten Hugel, Johannes Buchner, Matthias Rief
The heat-shock protein 90 (Hsp90) molecular chaperones are highly conserved across species. However, their dynamic properties can vary significantly from organism to organism. Here we used high-precision optical tweezers to analyze the mechanical properties and folding of different Hsp90 orthologs, namely bacterial Hsp90 (HtpG) and Hsp90 from the endoplasmic reticulum (ER) (Grp94), as well as from the cytosol of the eukaryotic cell (Hsp82). We find that the folding rates of Hsp82 and HtpG are similar, while the folding of Grp94 is slowed down by misfolding of the N-terminal domain. Furthermore, the domain interactions mediated by the charged linker, involved in the conformational cycles of all three orthologs, are much stronger for Grp94 than for Hsp82, keeping the N-terminal domain and the middle domain in close proximity. Thus, the ER resident Hsp90 ortholog differs from the cytosolic counterparts in basic functionally relevant structural properties.
DOI
The heat-shock protein 90 (Hsp90) molecular chaperones are highly conserved across species. However, their dynamic properties can vary significantly from organism to organism. Here we used high-precision optical tweezers to analyze the mechanical properties and folding of different Hsp90 orthologs, namely bacterial Hsp90 (HtpG) and Hsp90 from the endoplasmic reticulum (ER) (Grp94), as well as from the cytosol of the eukaryotic cell (Hsp82). We find that the folding rates of Hsp82 and HtpG are similar, while the folding of Grp94 is slowed down by misfolding of the N-terminal domain. Furthermore, the domain interactions mediated by the charged linker, involved in the conformational cycles of all three orthologs, are much stronger for Grp94 than for Hsp82, keeping the N-terminal domain and the middle domain in close proximity. Thus, the ER resident Hsp90 ortholog differs from the cytosolic counterparts in basic functionally relevant structural properties.
DOI
Optical Tweezers: Fundamentals and Some Biophysical Applications
Kamal R.Dhakal, Vasudevan Lakshminarayanan
Electromagnetic radiation changes its momentum when it interacts with small particles and a gradient force is experienced by the particle. This can be utilized to manipulate microscopic particles in an optical trap and is commonly referred to as optical tweezing. Optical tweezers are used as multifunctional tools in a myriad of applications such as micromanipulation, nanofabrication, biological studies of DNA, cells, biological micrometers, etc. This chapter will discuss the basic theory of optical tweezers (both wave theoretical and ray optics approximation) and provide examples from biomedical science and nanotechnology.
DOI
Electromagnetic radiation changes its momentum when it interacts with small particles and a gradient force is experienced by the particle. This can be utilized to manipulate microscopic particles in an optical trap and is commonly referred to as optical tweezing. Optical tweezers are used as multifunctional tools in a myriad of applications such as micromanipulation, nanofabrication, biological studies of DNA, cells, biological micrometers, etc. This chapter will discuss the basic theory of optical tweezers (both wave theoretical and ray optics approximation) and provide examples from biomedical science and nanotechnology.
DOI
Nanomaterials for in vivo imaging of mechanical forces and electrical fields
Randy D. Mehlenbacher, Rea Kolbl, Alice Lay & Jennifer A. Dionne
Cellular signalling is governed in large part by mechanical forces and electromagnetic fields. Mechanical forces play a critical role in cell differentiation, tissue organization and diseases such as cancer and heart disease; electrical fields are essential for intercellular communication, muscle contraction, neural signalling and sensory perception. Therefore, quantifying a biological system's forces and fields is crucial for understanding physiology and disease pathology and for developing medical tools for repair and recovery. This Review highlights advances in sensing mechanical forces and electrical fields in vivo, focusing on optical probes. The emergence of biocompatible optical probes, such as genetically encoded voltage indicators, molecular rotors, fluorescent dyes, semiconducting nanoparticles, plasmonic nanoparticles and lanthanide-doped upconverting nanoparticles, offers exciting opportunities to push the limits of spatial and temporal resolution, stability, multi-modality and stimuli sensitivity in bioimaging. We further discuss the materials design principles behind these probes and compare them across various metrics to facilitate sensor selection. Finally, we examine which advances are necessary to fully unravel the role of mechanical forces and electrical fields in vivo, such as the ability to probe the vectorial nature of forces, the development of combined force and field sensors, and the design of efficient optical actuators.
DOI
Cellular signalling is governed in large part by mechanical forces and electromagnetic fields. Mechanical forces play a critical role in cell differentiation, tissue organization and diseases such as cancer and heart disease; electrical fields are essential for intercellular communication, muscle contraction, neural signalling and sensory perception. Therefore, quantifying a biological system's forces and fields is crucial for understanding physiology and disease pathology and for developing medical tools for repair and recovery. This Review highlights advances in sensing mechanical forces and electrical fields in vivo, focusing on optical probes. The emergence of biocompatible optical probes, such as genetically encoded voltage indicators, molecular rotors, fluorescent dyes, semiconducting nanoparticles, plasmonic nanoparticles and lanthanide-doped upconverting nanoparticles, offers exciting opportunities to push the limits of spatial and temporal resolution, stability, multi-modality and stimuli sensitivity in bioimaging. We further discuss the materials design principles behind these probes and compare them across various metrics to facilitate sensor selection. Finally, we examine which advances are necessary to fully unravel the role of mechanical forces and electrical fields in vivo, such as the ability to probe the vectorial nature of forces, the development of combined force and field sensors, and the design of efficient optical actuators.
DOI
Optical orbital angular momentum: twisted light and chirality
Kayn A. Forbes and David L. Andrews
The question of how the orbital angular momentum of structured light might engage with chiral matter is a topic of resurgent interest. By taking account of electric quadrupole transition moments, it is shown that the handedness of the beam can indeed be exhibited in local chiral effects, being dependent on the sign of the topological charge. In the specific case of absorption, a significant interplay of wavefront structure and polarization is resolved, and clear differences in behavior are identified for systems possessing a degree of orientational order and for those that are randomly oriented.
DOI
The question of how the orbital angular momentum of structured light might engage with chiral matter is a topic of resurgent interest. By taking account of electric quadrupole transition moments, it is shown that the handedness of the beam can indeed be exhibited in local chiral effects, being dependent on the sign of the topological charge. In the specific case of absorption, a significant interplay of wavefront structure and polarization is resolved, and clear differences in behavior are identified for systems possessing a degree of orientational order and for those that are randomly oriented.
DOI
Tailoring Optical Gradient Force and Optical Scattering and Absorption Force
Junjie Du, Chi-Hong Yuen, Xiao Li, Kun Ding, Guiqiang Du, Zhifang Lin, C. T. Chan & Jack Ng
The introduction of the concept of gradient force and scattering and absorption force is an important milestone in optical trapping. However the profiles of these forces are usually unknown, even for standard setups. Here, we successfully calculated them analytically via multipole expansion and numerically via Mie theory and fast Fourier transform. The former provides physical insight, while the latter is highly accurate and efficient. A recipe to create truly conservative energy landscapes is presented. These may open up qualitatively new features in optical manipulation.
DOI
The introduction of the concept of gradient force and scattering and absorption force is an important milestone in optical trapping. However the profiles of these forces are usually unknown, even for standard setups. Here, we successfully calculated them analytically via multipole expansion and numerically via Mie theory and fast Fourier transform. The former provides physical insight, while the latter is highly accurate and efficient. A recipe to create truly conservative energy landscapes is presented. These may open up qualitatively new features in optical manipulation.
DOI
Finding trap stiffness of optical tweezers using digital filters
Pedro Almendarez-Rangel, Beatriz Morales-Cruzado, Erick Sarmiento-Gómez, and Francisco G. Pérez-Gutiérrez
Obtaining trap stiffness and calibration of the position detection system is the basis of a force measurement using optical tweezers. Both calibration quantities can be calculated using several experimental methods available in the literature. In most cases, stiffness determination and detection system calibration are performed separately, often requiring procedures in very different conditions, and thus confidence of calibration methods is not assured due to possible changes in the environment. In this work, a new method to simultaneously obtain both the detection system calibration and trap stiffness is presented. The method is based on the calculation of the power spectral density of positions through digital filters to obtain the harmonic contributions of the position signal. This method has the advantage of calculating both trap stiffness and photodetector calibration factor from the same dataset in situ. It also provides a direct method to avoid unwanted frequencies that could greatly affect calibration procedure, such as electric noise, for example.
DOI
Obtaining trap stiffness and calibration of the position detection system is the basis of a force measurement using optical tweezers. Both calibration quantities can be calculated using several experimental methods available in the literature. In most cases, stiffness determination and detection system calibration are performed separately, often requiring procedures in very different conditions, and thus confidence of calibration methods is not assured due to possible changes in the environment. In this work, a new method to simultaneously obtain both the detection system calibration and trap stiffness is presented. The method is based on the calculation of the power spectral density of positions through digital filters to obtain the harmonic contributions of the position signal. This method has the advantage of calculating both trap stiffness and photodetector calibration factor from the same dataset in situ. It also provides a direct method to avoid unwanted frequencies that could greatly affect calibration procedure, such as electric noise, for example.
DOI
Wednesday, January 24, 2018
Modeling and Control of Single-Cell Migration Induced by a Chemoattractant-Loaded Microbead
Ke Meng ; Hao Yang ; Yong Wang ; Dong Sun
Cell migration plays an essential role in cancer cell study. Investigation of a novel method for controlling cell migration movement can help develop new therapeutic strategies. In this paper, a chemoattractant-loaded microbead, which is controlled by optical tweezers, is used to stimulate a target cell to accomplish automated migration along a desired path while avoiding obstacles. Models of both tweezers-bead and bead-cell interactions are investigated. A dual closed-loop control strategy is proposed, which includes an inner tweezers-bead control loop and an outer bead-cell control loop. A proportional-integral feedback plus feedforward controller is used to control the inner loop, and an active disturbance rejection controller is used for the outer loop, which can address the cell migration modeling errors and unknown external disturbances. A traffic rule based on interference-clearing mechanism is also proposed to reduce external disturbances on the system by preventing other particles from interfering with the migration process. The effectiveness of the proposed control approach is verified by simulations and experiments on migrating leukemia cancer cells.
DOI
Cell migration plays an essential role in cancer cell study. Investigation of a novel method for controlling cell migration movement can help develop new therapeutic strategies. In this paper, a chemoattractant-loaded microbead, which is controlled by optical tweezers, is used to stimulate a target cell to accomplish automated migration along a desired path while avoiding obstacles. Models of both tweezers-bead and bead-cell interactions are investigated. A dual closed-loop control strategy is proposed, which includes an inner tweezers-bead control loop and an outer bead-cell control loop. A proportional-integral feedback plus feedforward controller is used to control the inner loop, and an active disturbance rejection controller is used for the outer loop, which can address the cell migration modeling errors and unknown external disturbances. A traffic rule based on interference-clearing mechanism is also proposed to reduce external disturbances on the system by preventing other particles from interfering with the migration process. The effectiveness of the proposed control approach is verified by simulations and experiments on migrating leukemia cancer cells.
DOI
Optical torque on small chiral particles in generic optical fields
Huajin Chen, Wanli Lu, Xinning Yu, Chunhua Xue, Shiyang Liu, and Zhifang Lin
We derive an analytical expression of the optical torque (OT) on chiral particles in generic monochromatic optical fields within the dipole approximation. Besides the extinction terms owing to the interaction between the dipoles excited on the particle and the incident field, our expression includes also some long missing terms that are understood as recoil due to the radiation of the dipoles excited on the particle. The recoil terms are shown to have a significant contribution to the OT in many situations. Inspired by our expression, we further proved that the net OT vanishes for a lossless isotropic chiral spherical particle of any size illuminated by arbitrary monochromatic optical fields. Finally, we trace the origin of OT on a small chiral sphere immersed in a zeroth-order vector Bessel beam, taking advantage of our analytical expression. It is found that the azimuthal OT perpendicular to the beam’s propagation direction comes from the transfer of the spin angular momentum of the incident field to the particle, while the longitudinal OT along the illumination direction originates from the particle chirality, which generates longitudinal angular momentum on the optical beam and thus makes the particle subject to a longitudinal OT by recoil. Thus the longitudinal OT tends to rotate the chiral particle with opposite helicities in opposite directions, while the transverse OT shows little dependence on the handedness of particle chirality. Our results may help in understanding OT as an extra handle for particle manipulations in optical tweezers.
DOI
We derive an analytical expression of the optical torque (OT) on chiral particles in generic monochromatic optical fields within the dipole approximation. Besides the extinction terms owing to the interaction between the dipoles excited on the particle and the incident field, our expression includes also some long missing terms that are understood as recoil due to the radiation of the dipoles excited on the particle. The recoil terms are shown to have a significant contribution to the OT in many situations. Inspired by our expression, we further proved that the net OT vanishes for a lossless isotropic chiral spherical particle of any size illuminated by arbitrary monochromatic optical fields. Finally, we trace the origin of OT on a small chiral sphere immersed in a zeroth-order vector Bessel beam, taking advantage of our analytical expression. It is found that the azimuthal OT perpendicular to the beam’s propagation direction comes from the transfer of the spin angular momentum of the incident field to the particle, while the longitudinal OT along the illumination direction originates from the particle chirality, which generates longitudinal angular momentum on the optical beam and thus makes the particle subject to a longitudinal OT by recoil. Thus the longitudinal OT tends to rotate the chiral particle with opposite helicities in opposite directions, while the transverse OT shows little dependence on the handedness of particle chirality. Our results may help in understanding OT as an extra handle for particle manipulations in optical tweezers.
DOI
Rotating of low-refractive-index microparticles with a quasi-perfect optical vortex
Yansheng Liang, Ming Lei, Shaohui Yan, Manman Li, Yanan Cai, Zhaojun Wang, Xianghua Yu, and Baoli Yao
Low-refractive-index microparticles, such as hollow microspheres, have shown great significance in some applications, such as biomedical sensing and targeted drug delivery. However, optical trapping and manipulation of low-refractive-index microparticles are challenging, owing to the repelling force exerted by typical optical traps. In this paper, we demonstrated optical trapping and rotating of large-sized low-refractive-index microparticles by using quasi-perfect optical vortex (quasi-POV) beams, which were generated by Fourier transform of high-order quasi-Bessel beams. Numerical simulation was carried out to characterize the focusing property of the quasi-POV beams. The dynamics of low-refractive-index microparticles in the quasi-POV with various topological charges was investigated in detail. To improve the trapping and rotating performances of the vortex, a point trap was introduced at the center of the ring. Experimental results showed that the quasi-POV was preferable for manipulation of large-sized low-refractive-index microparticles, with its control of the particles’ rotating velocity dependent only on the topological charge due to the unchanged orbital radius.
DOI
Low-refractive-index microparticles, such as hollow microspheres, have shown great significance in some applications, such as biomedical sensing and targeted drug delivery. However, optical trapping and manipulation of low-refractive-index microparticles are challenging, owing to the repelling force exerted by typical optical traps. In this paper, we demonstrated optical trapping and rotating of large-sized low-refractive-index microparticles by using quasi-perfect optical vortex (quasi-POV) beams, which were generated by Fourier transform of high-order quasi-Bessel beams. Numerical simulation was carried out to characterize the focusing property of the quasi-POV beams. The dynamics of low-refractive-index microparticles in the quasi-POV with various topological charges was investigated in detail. To improve the trapping and rotating performances of the vortex, a point trap was introduced at the center of the ring. Experimental results showed that the quasi-POV was preferable for manipulation of large-sized low-refractive-index microparticles, with its control of the particles’ rotating velocity dependent only on the topological charge due to the unchanged orbital radius.
DOI
Effect of Bovine Serum Albumin on Red Blood Cell Optical Anisotropy Probed Through the Optomechanical Response in an Optical Trap
Parthasarathi Praveen, Selvan Rekha, Devarkonda Chetana, Shruthi S. Iyengar, Sarbari Bhattacharya, Sharath Ananthamu
The dynamics of trapped entities in an Optical Trap (OT) can yield information with regards to their viscoelastic response as well as optical anisotropy, if any. Detailed analysis of such dynamics correlated with parameters which affect the response can yield additional clues to the exact effect of these on the trapped entities. In this work, we illustrate this point by showing how the altered behavior of Red Blood Cells (RBC) treated with Bovine Serum Albumin (BSA) yields information about the nature of action of BSA, on which there is no current consensus in literature. We conclude from our studies that BSA treatment leads to a change in the birefringence of the RBCs, a conclusion arrived at from the altered optomechanical response of such cells in a linearly polarized Gaussian beam OT. Furthermore, we argue that the observed changes in cellular optical anisotropy may be thought of as due to changes in the curvature of the RBC membrane. We also note that BSA action could help mimic pathological conditions that result in an altered cell shape.
DOI
The dynamics of trapped entities in an Optical Trap (OT) can yield information with regards to their viscoelastic response as well as optical anisotropy, if any. Detailed analysis of such dynamics correlated with parameters which affect the response can yield additional clues to the exact effect of these on the trapped entities. In this work, we illustrate this point by showing how the altered behavior of Red Blood Cells (RBC) treated with Bovine Serum Albumin (BSA) yields information about the nature of action of BSA, on which there is no current consensus in literature. We conclude from our studies that BSA treatment leads to a change in the birefringence of the RBCs, a conclusion arrived at from the altered optomechanical response of such cells in a linearly polarized Gaussian beam OT. Furthermore, we argue that the observed changes in cellular optical anisotropy may be thought of as due to changes in the curvature of the RBC membrane. We also note that BSA action could help mimic pathological conditions that result in an altered cell shape.
DOI
Multiplexed fluctuation-dissipation-theorem calibration of optical tweezers inside living cells
Hao Yan, Jessica F. Johnston, Sidney B. Cahn, Megan C. King, and Simon G. J. Mochrie
In order to apply optical tweezers-based force measurements within an uncharacterized viscoelastic medium such as the cytoplasm of a living cell, a quantitative calibration method that may be applied in this complex environment is needed. We describe an improved version of the fluctuation-dissipation-theorem calibration method, which has been developed to perform in situ calibration in viscoelastic media without prior knowledge of the trapped object. Using this calibration procedure, it is possible to extract values of the medium’s viscoelastic moduli as well as the force constant describing the optical trap. To demonstrate our method, we calibrate an optical trap in water, in polyethylene oxide solutions of different concentrations, and inside living fission yeast (S. pombe).
DOI
In order to apply optical tweezers-based force measurements within an uncharacterized viscoelastic medium such as the cytoplasm of a living cell, a quantitative calibration method that may be applied in this complex environment is needed. We describe an improved version of the fluctuation-dissipation-theorem calibration method, which has been developed to perform in situ calibration in viscoelastic media without prior knowledge of the trapped object. Using this calibration procedure, it is possible to extract values of the medium’s viscoelastic moduli as well as the force constant describing the optical trap. To demonstrate our method, we calibrate an optical trap in water, in polyethylene oxide solutions of different concentrations, and inside living fission yeast (S. pombe).
DOI
Enhancement of the ‘tractor-beam’ pulling force on an optically bound structure
Jana Damková, Lukáš Chvátal, Jan Ježek, Jindřich Oulehla, Oto Brzobohatý & Pavel Zemánek
Recently, increasing attention has been devoted to mastering a new technique of optical delivery of micro-objects tractor-beam’. Such beams have uniform intensity profiles along their propagation direction and can exert a negative force that, in contrast to the familiar pushing force associated with radiation pressure, pulls the scatterer toward the light source. It was experimentally observed that under certain circumstances, the pulling force can be significantly enhanced if a non-spherical scatterer, for example, a linear chain of optically bound objects, is optically transported. Here we demonstrate that motion of two optically bound objects in a tractor beam strongly depends on theirs mutual distance and spatial orientation. Such configuration-dependent optical forces add extra flexibility to our ability to control matter with light. Understanding these interactions opens the door to new applications involving the formation, sorting or delivery of colloidal self-organized structures.
DOI
Recently, increasing attention has been devoted to mastering a new technique of optical delivery of micro-objects tractor-beam’. Such beams have uniform intensity profiles along their propagation direction and can exert a negative force that, in contrast to the familiar pushing force associated with radiation pressure, pulls the scatterer toward the light source. It was experimentally observed that under certain circumstances, the pulling force can be significantly enhanced if a non-spherical scatterer, for example, a linear chain of optically bound objects, is optically transported. Here we demonstrate that motion of two optically bound objects in a tractor beam strongly depends on theirs mutual distance and spatial orientation. Such configuration-dependent optical forces add extra flexibility to our ability to control matter with light. Understanding these interactions opens the door to new applications involving the formation, sorting or delivery of colloidal self-organized structures.
DOI
Tuesday, January 23, 2018
Microfluidics: A new tool for microbial single cell analyses in human microbiome studies
Yuguang Liu and Marina Walther-Antonio
Microbial cells behave differently in colonies and when singled out. The standard methods of microbiome studies present the average characteristics and behaviors of heterogeneous populations and lack the resolution to analyze microbes on a single cell level. Besides, the microbiome does not exist in pure colonies in human bodies, but naturally in complex communities. Therefore, it would be ideal to observe the microbes on a single cell level while maintaining their natural settings. Conventional dilution-to-extinction methods are capable of reducing the complexity of the communities to a minimal ecologically functional unit; however, new tools are needed to perform these studies. Recently, microfluidics as a new technology is gaining attention for various single cell applications because it offers the unique ability of handling nanoscale volume in microstructures, providing an attractive alternative to look at single microbial cells. Here, we compare different microfluidic technologies for microbial single cell studies and review the advancement of microfluidics for various microbial single cell analyses. Continuous microfluidics has been used in microbial single cell culture, whole genome sequencing, gene expression, and metabolic analyses; however, droplet microfluidics is becoming a new trend for its high-throughput single cell encapsulation ability. We envision that different microfluidic paradigms will be integrated in the future for their unique attributes and offer a versatile platform for systematic microbiome studies.
DOI
Microbial cells behave differently in colonies and when singled out. The standard methods of microbiome studies present the average characteristics and behaviors of heterogeneous populations and lack the resolution to analyze microbes on a single cell level. Besides, the microbiome does not exist in pure colonies in human bodies, but naturally in complex communities. Therefore, it would be ideal to observe the microbes on a single cell level while maintaining their natural settings. Conventional dilution-to-extinction methods are capable of reducing the complexity of the communities to a minimal ecologically functional unit; however, new tools are needed to perform these studies. Recently, microfluidics as a new technology is gaining attention for various single cell applications because it offers the unique ability of handling nanoscale volume in microstructures, providing an attractive alternative to look at single microbial cells. Here, we compare different microfluidic technologies for microbial single cell studies and review the advancement of microfluidics for various microbial single cell analyses. Continuous microfluidics has been used in microbial single cell culture, whole genome sequencing, gene expression, and metabolic analyses; however, droplet microfluidics is becoming a new trend for its high-throughput single cell encapsulation ability. We envision that different microfluidic paradigms will be integrated in the future for their unique attributes and offer a versatile platform for systematic microbiome studies.
DOI
Insight into plasmonic hot-electron transfer and plasmon molecular drive: new dimensions in energy conversion and nanofabrication
Akihiro Furube & Shuichi Hashimoto
Localized surface plasmon resonance (LSPR) of plasmonic nanoparticles and nanostructures has attracted wide attention because the nanoparticles exhibit a strong near-field enhancement through interaction with visible light, enabling subwavelength optics and sensing at the single-molecule level. The extremely fast LSPR decays have raised doubts that such nanoparticles have use in photochemistry and energy storage. Recent studies have demonstrated the capability of such plasmonic systems in producing LSPR-induced hot electrons that are useful in energy conversion and storage when combined with electron-accepting semiconductors. Due to the femtosecond timescale, hot-electron transfer is under intense investigation to promote ongoing applications in photovoltaics and photocatalysis. Concurrently, hot-electron decay results in photothermal responses or plasmonic heating. Importantly, this heating has received renewed interest in photothermal manipulation, despite the developments in optical manipulation using optical forces to move and position nanoparticles and molecules guided by plasmonic nanostructures. To realize plasmonic heating-based manipulation, photothermally generated flows, such as thermophoresis, the Marangoni effect and thermal convection, are exploited. Plasmon-enhanced optical tweezers together with plasmon-induced heating show potential as an ultimate bottom-up method for fabricating nanomaterials. We review recent progress in two fascinating areas: solar energy conversion through interfacial electron transfer in gold-semiconductor composite materials and plasmon-induced nanofabrication.
DOI
Localized surface plasmon resonance (LSPR) of plasmonic nanoparticles and nanostructures has attracted wide attention because the nanoparticles exhibit a strong near-field enhancement through interaction with visible light, enabling subwavelength optics and sensing at the single-molecule level. The extremely fast LSPR decays have raised doubts that such nanoparticles have use in photochemistry and energy storage. Recent studies have demonstrated the capability of such plasmonic systems in producing LSPR-induced hot electrons that are useful in energy conversion and storage when combined with electron-accepting semiconductors. Due to the femtosecond timescale, hot-electron transfer is under intense investigation to promote ongoing applications in photovoltaics and photocatalysis. Concurrently, hot-electron decay results in photothermal responses or plasmonic heating. Importantly, this heating has received renewed interest in photothermal manipulation, despite the developments in optical manipulation using optical forces to move and position nanoparticles and molecules guided by plasmonic nanostructures. To realize plasmonic heating-based manipulation, photothermally generated flows, such as thermophoresis, the Marangoni effect and thermal convection, are exploited. Plasmon-enhanced optical tweezers together with plasmon-induced heating show potential as an ultimate bottom-up method for fabricating nanomaterials. We review recent progress in two fascinating areas: solar energy conversion through interfacial electron transfer in gold-semiconductor composite materials and plasmon-induced nanofabrication.
DOI
A change of view: homologous recombination at single-molecule resolution
Kyle Kaniecki, Luisina De Tullio & Eric C. Greene
Genetic recombination occurs in all organisms and is vital for genome stability. Indeed, in humans, aberrant recombination can lead to diseases such as cancer. Our understanding of homologous recombination is built upon more than a century of scientific inquiry, but achieving a more complete picture using ensemble biochemical and genetic approaches is hampered by population heterogeneity and transient recombination intermediates. Recent advances in single-molecule and super-resolution microscopy methods help to overcome these limitations and have led to new and refined insights into recombination mechanisms, including a detailed understanding of DNA helicase function and synaptonemal complex structure. The ability to view cellular processes at single-molecule resolution promises to transform our understanding of recombination and related processes.
DOI
Genetic recombination occurs in all organisms and is vital for genome stability. Indeed, in humans, aberrant recombination can lead to diseases such as cancer. Our understanding of homologous recombination is built upon more than a century of scientific inquiry, but achieving a more complete picture using ensemble biochemical and genetic approaches is hampered by population heterogeneity and transient recombination intermediates. Recent advances in single-molecule and super-resolution microscopy methods help to overcome these limitations and have led to new and refined insights into recombination mechanisms, including a detailed understanding of DNA helicase function and synaptonemal complex structure. The ability to view cellular processes at single-molecule resolution promises to transform our understanding of recombination and related processes.
DOI
Stochastic control for optical manipulation of multiple microscopic objects
Quang Minh Ta, Chien Chern Cheah
While various control techniques have been developed for optical manipulation, the Brownian movement of microscopic objects in the medium is usually ignored for simplicity of analyzing the control systems. Nevertheless, due to the universality of the Brownian movement and its effect on optical manipulation of cells or micro-objects, it is required for the Brownian effect to be properly taken into consideration so as to ensure the stability and performance of the control systems. In this paper, we derive a stochastic control technique to achieve a theoretical framework for optical manipulation of multiple microscopic objects in the presence of the Brownian perturbations. In the proposed control methodology, a region control technique and a dynamic interaction approach are developed for collision-free manipulation of the target micro-objects with random perturbations. All the target micro-objects are trapped and manipulated simultaneously while being kept inside the desired dynamic region, and at the same time, preserving a minimum distance with each other to avoid collisions. While a bounded tracking or region error exists in current control techniques for optical manipulation due to the effect of the Brownian perturbations, this paper provides a new approach which guarantees that all the target micro-objects are kept inside the desired region during the course of manipulation. Rigorous mathematical formulation has been developed for automated manipulation of multiple microscopic objects in the presence of the Brownian perturbations, and experimental results are presented to demonstrate the feasibility and effectiveness of the proposed control technique.
DOI
While various control techniques have been developed for optical manipulation, the Brownian movement of microscopic objects in the medium is usually ignored for simplicity of analyzing the control systems. Nevertheless, due to the universality of the Brownian movement and its effect on optical manipulation of cells or micro-objects, it is required for the Brownian effect to be properly taken into consideration so as to ensure the stability and performance of the control systems. In this paper, we derive a stochastic control technique to achieve a theoretical framework for optical manipulation of multiple microscopic objects in the presence of the Brownian perturbations. In the proposed control methodology, a region control technique and a dynamic interaction approach are developed for collision-free manipulation of the target micro-objects with random perturbations. All the target micro-objects are trapped and manipulated simultaneously while being kept inside the desired dynamic region, and at the same time, preserving a minimum distance with each other to avoid collisions. While a bounded tracking or region error exists in current control techniques for optical manipulation due to the effect of the Brownian perturbations, this paper provides a new approach which guarantees that all the target micro-objects are kept inside the desired region during the course of manipulation. Rigorous mathematical formulation has been developed for automated manipulation of multiple microscopic objects in the presence of the Brownian perturbations, and experimental results are presented to demonstrate the feasibility and effectiveness of the proposed control technique.
DOI
Spinning of particles in optical double-vortex beams
Manman Li, Shaohui Yan, Yansheng Liang, Peng Zhang and Baoli Yao
Optical spin angular momentum, an intrinsic part of optical angular momemtum, can induce a spinning motion of a trapped particle around its own axis in optical manipulation. Focusing of a type of double-vortex (DV) input field obtained by linearly superposing two optical vortex beams with equal but opposite topological charges, yields a multi-lobe focal field, each of which has non-vanishing optical spin angular momentum, and is capable of trapping particle while spinning the particle around a certain axis. Significantly, both the focusing properties and the spinning dynamics are strongly polarization dependent. For instance, the focused field of a circularly polarized double-vortex (CPDV) beam carries transverse and longitudinal spin angular momenta, inducing axial spinning of the trapped particles, whereas the focused field of a radially polarized double-vortex (RPDV) beam possesses purely transverse spin angular momentum and can drive the particles to spin transversely to the optical axis. These results may find potential applications in light beam shaping and optical manipulations.
DOI
Optical spin angular momentum, an intrinsic part of optical angular momemtum, can induce a spinning motion of a trapped particle around its own axis in optical manipulation. Focusing of a type of double-vortex (DV) input field obtained by linearly superposing two optical vortex beams with equal but opposite topological charges, yields a multi-lobe focal field, each of which has non-vanishing optical spin angular momentum, and is capable of trapping particle while spinning the particle around a certain axis. Significantly, both the focusing properties and the spinning dynamics are strongly polarization dependent. For instance, the focused field of a circularly polarized double-vortex (CPDV) beam carries transverse and longitudinal spin angular momenta, inducing axial spinning of the trapped particles, whereas the focused field of a radially polarized double-vortex (RPDV) beam possesses purely transverse spin angular momentum and can drive the particles to spin transversely to the optical axis. These results may find potential applications in light beam shaping and optical manipulations.
DOI
Heterogeneous Capillary Interactions of Interface-Trapped Ellipsoid Particles Using the Trap-Release Method
Jin Hyun Lim, Jun Young Kim, Dong Woo Kang, Kyu Hwan Choi, Seong Jae Lee, Sang Hyuk Im, and Bum Jun Park
Heterogeneous capillary interactions between ellipsoid particles at the oil–water interface were measured via optical laser tweezers. Two trapped particles were aligned in either tip-to-tip (tt) or side-to-side (ss) configurations via the double-trap method and were released from the optical traps, leading to particle–particle attractions due to the capillary forces caused by quadrupolar interface deformation. On the basis of image analysis and calculations of the Stokes drag force, the capillary interactions between two ellipsoid particles with the same aspect ratio (E) were found to vary with the particle pairs that were measured, indicating that the interactions were nondeterministic or heterogeneous. Heterogeneous capillary interactions could be attributed to undulation of the interface meniscus due to chemical and/or geometric particle heterogeneity. The power law exponent for the capillary interaction Ucap ≈ r–β was found to be β ≈ 4 and was independent of the aspect ratio and particle configuration in long-range separations. Additionally, with regard to the tt configuration, the magnitude of the capillary force proportionally increased with the E value (E > 1) when two ellipsoid particles approached each other in the tt configuration.
DOI
Heterogeneous capillary interactions between ellipsoid particles at the oil–water interface were measured via optical laser tweezers. Two trapped particles were aligned in either tip-to-tip (tt) or side-to-side (ss) configurations via the double-trap method and were released from the optical traps, leading to particle–particle attractions due to the capillary forces caused by quadrupolar interface deformation. On the basis of image analysis and calculations of the Stokes drag force, the capillary interactions between two ellipsoid particles with the same aspect ratio (E) were found to vary with the particle pairs that were measured, indicating that the interactions were nondeterministic or heterogeneous. Heterogeneous capillary interactions could be attributed to undulation of the interface meniscus due to chemical and/or geometric particle heterogeneity. The power law exponent for the capillary interaction Ucap ≈ r–β was found to be β ≈ 4 and was independent of the aspect ratio and particle configuration in long-range separations. Additionally, with regard to the tt configuration, the magnitude of the capillary force proportionally increased with the E value (E > 1) when two ellipsoid particles approached each other in the tt configuration.
DOI
Monday, January 22, 2018
Design of an optical conveyor for selective separation of a mixture of enantiomers
P. Acebal, L. Carretero, and S. Blaya
Chiral resolution is a fundamental problem in pharmaceutics or agrochemicals, so a great effort has been made to generate experimental techniques capable of producing mechanical separation of a mixture of enantiomers. Unlike other techniques that are usually employed, such as chiral resolving agents or chiral chromatography, we propose a new technique which is directly applicable in solution and without further processing. This technique is based on optical forces, since we show that with the proper design of the polarization states of the incident beams and temporal dephasing, a chiral sensitive optical conveyor can be obtained that is able to transport enantiomers in opposite directions. The implementation of such an optical conveyor with the required focused optical fields produces a well-defined trapping region for each enantiomer, since theoretical simulations over a large number of chiral particle trajectories show that it is possible to reach values of enantiomeric excess of over 99%.
DOI
Chiral resolution is a fundamental problem in pharmaceutics or agrochemicals, so a great effort has been made to generate experimental techniques capable of producing mechanical separation of a mixture of enantiomers. Unlike other techniques that are usually employed, such as chiral resolving agents or chiral chromatography, we propose a new technique which is directly applicable in solution and without further processing. This technique is based on optical forces, since we show that with the proper design of the polarization states of the incident beams and temporal dephasing, a chiral sensitive optical conveyor can be obtained that is able to transport enantiomers in opposite directions. The implementation of such an optical conveyor with the required focused optical fields produces a well-defined trapping region for each enantiomer, since theoretical simulations over a large number of chiral particle trajectories show that it is possible to reach values of enantiomeric excess of over 99%.
DOI
Single Actin Bundle Rheology
Dan Strehle, Paul Mollenkopf, Martin Glaser, Tom Golde, Carsten Schuldt, Josef A. Käs and Jörg Schnauß
Bundled actin structures play an essential role in the mechanical response of the actin cytoskeleton in eukaryotic cells. Although responsible for crucial cellular processes, they are rarely investigated in comparison to single filaments and isotropic networks. Presenting a highly anisotropic structure, the determination of the mechanical properties of individual bundles was previously achieved through passive approaches observing bending deformations induced by thermal fluctuations. We present a new method to determine the bending stiffness of individual bundles, by measuring the decay of an actively induced oscillation. This approach allows us to systematically test anisotropic, bundled structures. Our experiments revealed that thin, depletion force-induced bundles behave as semiflexible polymers and obey the theoretical predictions determined by the wormlike chain model. Thickening an individual bundle by merging it with other bundles enabled us to study effects that are solely based on the number of involved filaments. These thicker bundles showed a frequency-dependent bending stiffness, a behavior that is inconsistent with the predictions of the wormlike chain model. We attribute this effect to internal processes and give a possible explanation with regard to the wormlike bundle theory.
DOI
Bundled actin structures play an essential role in the mechanical response of the actin cytoskeleton in eukaryotic cells. Although responsible for crucial cellular processes, they are rarely investigated in comparison to single filaments and isotropic networks. Presenting a highly anisotropic structure, the determination of the mechanical properties of individual bundles was previously achieved through passive approaches observing bending deformations induced by thermal fluctuations. We present a new method to determine the bending stiffness of individual bundles, by measuring the decay of an actively induced oscillation. This approach allows us to systematically test anisotropic, bundled structures. Our experiments revealed that thin, depletion force-induced bundles behave as semiflexible polymers and obey the theoretical predictions determined by the wormlike chain model. Thickening an individual bundle by merging it with other bundles enabled us to study effects that are solely based on the number of involved filaments. These thicker bundles showed a frequency-dependent bending stiffness, a behavior that is inconsistent with the predictions of the wormlike chain model. We attribute this effect to internal processes and give a possible explanation with regard to the wormlike bundle theory.
DOI
Techniques to stimulate and interrogate cell-cell adhesion mechanics
Ruiguo Yang, Joshua A. Broussard, Kathleen J. Green, Horacio D. Espinosa
Cell–cell adhesions maintain the mechanical integrity of multicellular tissues and have recently been found to act as mechanotransducers, translating mechanical cues into biochemical signals. Mechanotransduction studies have primarily focused on focal adhesions, sites of cell-substrate attachment. These studies leverage technical advances in devices and systems interfacing with living cells through cell-extracellular matrix adhesions. As reports of aberrant signal transduction originating from mutations in cell–cell adhesion molecules are being increasingly associated with disease states, growing attention is being paid to this intercellular signaling hub. Along with this renewed focus, new requirements arise for the interrogation and stimulation of cell–cell adhesive junctions. This review covers established experimental techniques for stimulation and interrogation of cell–cell adhesion from cell pairs to monolayers.
DOI
Cell–cell adhesions maintain the mechanical integrity of multicellular tissues and have recently been found to act as mechanotransducers, translating mechanical cues into biochemical signals. Mechanotransduction studies have primarily focused on focal adhesions, sites of cell-substrate attachment. These studies leverage technical advances in devices and systems interfacing with living cells through cell-extracellular matrix adhesions. As reports of aberrant signal transduction originating from mutations in cell–cell adhesion molecules are being increasingly associated with disease states, growing attention is being paid to this intercellular signaling hub. Along with this renewed focus, new requirements arise for the interrogation and stimulation of cell–cell adhesive junctions. This review covers established experimental techniques for stimulation and interrogation of cell–cell adhesion from cell pairs to monolayers.
DOI
Mfd Dynamically Regulates Transcription via a Release and Catch-Up Mechanism
Tung T. Le, Yi Yang, Chuang Tan, Margaret M. Suhanovsky, Robert M. Fulbright Jr., James T. Inman, Ming Li, Jaeyoon Lee, Sarah Perelman, Jeffrey W. Roberts, Alexandra M. Deaconescu, Michelle D. Wang
The bacterial Mfd ATPase is increasingly recognized as a general transcription factor that participates in the resolution of transcription conflicts with other processes/roadblocks. This function stems from Mfd’s ability to preferentially act on stalled RNA polymerases (RNAPs). However, the mechanism underlying this preference and the subsequent coordination between Mfd and RNAP have remained elusive. Here, using a novel real-time translocase assay, we unexpectedly discovered that Mfd translocates autonomously on DNA. The speed and processivity of Mfd dictate a “release and catch-up” mechanism to efficiently patrol DNA for frequently stalled RNAPs. Furthermore, we showed that Mfd prevents RNAP backtracking or rescues a severely backtracked RNAP, allowing RNAP to overcome stronger obstacles. However, if an obstacle’s resistance is excessive, Mfd dissociates the RNAP, clearing the DNA for other processes. These findings demonstrate a remarkably delicate coordination between Mfd and RNAP, allowing efficient targeting and recycling of Mfd and expedient conflict resolution.
DOI
The bacterial Mfd ATPase is increasingly recognized as a general transcription factor that participates in the resolution of transcription conflicts with other processes/roadblocks. This function stems from Mfd’s ability to preferentially act on stalled RNA polymerases (RNAPs). However, the mechanism underlying this preference and the subsequent coordination between Mfd and RNAP have remained elusive. Here, using a novel real-time translocase assay, we unexpectedly discovered that Mfd translocates autonomously on DNA. The speed and processivity of Mfd dictate a “release and catch-up” mechanism to efficiently patrol DNA for frequently stalled RNAPs. Furthermore, we showed that Mfd prevents RNAP backtracking or rescues a severely backtracked RNAP, allowing RNAP to overcome stronger obstacles. However, if an obstacle’s resistance is excessive, Mfd dissociates the RNAP, clearing the DNA for other processes. These findings demonstrate a remarkably delicate coordination between Mfd and RNAP, allowing efficient targeting and recycling of Mfd and expedient conflict resolution.
DOI
Single-molecule measurements of the effect of force on Thy-1/αvβ3-integrin interaction using non-purified proteins
Francesca Burgos-Bravo, Nataniel L. Figueroa, Nathalie Casanova-Morales, Andrew F. G. Quest, Christian A. M. Wilson, and Lisette Leyton
Thy-1 and αvβ3 integrin mediate bidirectional cell-to-cell communication between neurons and astrocytes. Thy-1/αvβ3 interactions stimulate astrocyte migration and the retraction of neuronal prolongations, both processes in which internal forces are generated affecting the bimolecular interactions that maintain cell-cell adhesion. Nonetheless, how the Thy-1/αvβ3 interactions respond to mechanical cues is an unresolved issue. In this study, optical tweezers were used as a single-molecule force transducer, and the Dudko-Hummer-Szabo Model was applied to calculate the kinetic parameters of Thy-1/αvβ3 dissociation. A novel experimental strategy was implemented to analyze the interaction of Thy-1-Fc with non-purified αvβ3-Fc integrin, whereby non-specific rupture events were corrected by using a new mathematical approach. This methodology permitted accurately estimating specific rupture forces for Thy-1-Fc/αvβ3-Fc dissociation and calculating the kinetic and transition state parameters. Force exponentially accelerated Thy-1/αvβ3 dissociation, indicating slip bond behavior. Importantly, non-specific interactions were detected even for purified proteins, highlighting the importance of correcting for such interactions. In conclusion, we describe a new strategy to characterize the response of bimolecular interactions to forces even in the presence of non-specific binding events. By defining how force regulates Thy-1/αvβ3 integrin binding, we provide an initial step towards understanding how the neuron-astrocyte pair senses and responds to mechanical cues.
DOI
Thy-1 and αvβ3 integrin mediate bidirectional cell-to-cell communication between neurons and astrocytes. Thy-1/αvβ3 interactions stimulate astrocyte migration and the retraction of neuronal prolongations, both processes in which internal forces are generated affecting the bimolecular interactions that maintain cell-cell adhesion. Nonetheless, how the Thy-1/αvβ3 interactions respond to mechanical cues is an unresolved issue. In this study, optical tweezers were used as a single-molecule force transducer, and the Dudko-Hummer-Szabo Model was applied to calculate the kinetic parameters of Thy-1/αvβ3 dissociation. A novel experimental strategy was implemented to analyze the interaction of Thy-1-Fc with non-purified αvβ3-Fc integrin, whereby non-specific rupture events were corrected by using a new mathematical approach. This methodology permitted accurately estimating specific rupture forces for Thy-1-Fc/αvβ3-Fc dissociation and calculating the kinetic and transition state parameters. Force exponentially accelerated Thy-1/αvβ3 dissociation, indicating slip bond behavior. Importantly, non-specific interactions were detected even for purified proteins, highlighting the importance of correcting for such interactions. In conclusion, we describe a new strategy to characterize the response of bimolecular interactions to forces even in the presence of non-specific binding events. By defining how force regulates Thy-1/αvβ3 integrin binding, we provide an initial step towards understanding how the neuron-astrocyte pair senses and responds to mechanical cues.
DOI
Study of non-covalent interactions on dendriplex formation: Influence of hydrophobic, electrostatic and hydrogen bonds interactions
María Sánchez-Milla, Isabel Pastor, Marek Maly, M. Jesús Serramía, Rafael Gómez, Javier Sánchez-Nieves, Félix Ritort, M. Ángeles Muñoz-Fernández, F. Javierde la Mata
The interaction of a double stranded small interference RNA (siRNA Nef) with cationic carbosilane dendrimers of generations 1–3 with two different ammonium functions at the periphery ([−NMe2R]+, R = Me, (CH2)2OH) has been studied by experimental techniques (zeta potential, electrophoresis, single molecule pulling experiments) and molecular dynamic calculations. These studies state the presence of different forces on dendriplex formation, depending on generation and type of ammonium group. Whilst for higher dendrimers electrostatic forces mainly drive the stability of dendriplexes, first generation compounds can penetrate into siRNA strands due to the establishment of hydrophobic interactions. Finally, in the particular case of first generation dendrimer [G1O3(NMe2(CH2)2OH))6]6+; the presence of hydroxyl groups reinforces dendriplex stability by hydrogen bonds formation. However, since these small dendrimers do not cover the RNA, only higher generation derivatives protect RNA from degradation.
DOI
The interaction of a double stranded small interference RNA (siRNA Nef) with cationic carbosilane dendrimers of generations 1–3 with two different ammonium functions at the periphery ([−NMe2R]+, R = Me, (CH2)2OH) has been studied by experimental techniques (zeta potential, electrophoresis, single molecule pulling experiments) and molecular dynamic calculations. These studies state the presence of different forces on dendriplex formation, depending on generation and type of ammonium group. Whilst for higher dendrimers electrostatic forces mainly drive the stability of dendriplexes, first generation compounds can penetrate into siRNA strands due to the establishment of hydrophobic interactions. Finally, in the particular case of first generation dendrimer [G1O3(NMe2(CH2)2OH))6]6+; the presence of hydroxyl groups reinforces dendriplex stability by hydrogen bonds formation. However, since these small dendrimers do not cover the RNA, only higher generation derivatives protect RNA from degradation.
DOI
Friday, January 19, 2018
Micro/nanofluidics-enabled single-cell biochemical analysis
Ling Lin, Qinghua Chen, Jiashu Sun
The micro/nanofluidic technique has become an important tool for single-cell analysis with the capability to integrate time-consuming and labour-intensive experimental procedures into a small device. Micro/nanofluidics-based biochemical analysis exhibits advantages over conventional methods in terms of small sample volume, rapid turnaround time, straightforward operation, and efficient processing. Single-cell manipulation, therapy, detection and sequencing could be implemented within a sophisticated and multi-functional micro/nanofluidic platform. Here we review recent developments of micro/nanofluidic technologies for single-cell analysis, with emphasis on cell trapping, treatment, and biochemical studies. The potential of micro/nanofluidics-based single-cell analysis is discussed.
DOI
DOI
Extended depth of field for single biomolecule optical imaging-force spectroscopy
Minhyeok Chang, Jungsic Oh, Yeonghoon Kim, Sungchul Hohng, and Jong-Bong Lee
Real-time optical imaging combined with single-molecule manipulation broadens the horizons for acquiring information about the spatiotemporal localization and the mechanical details of target molecules. To obtain an optical signal outside the focal plane without unintended interruption of the force signal in single-molecule optical imaging-force spectroscopy, we developed an optical method to extend the depth of field in a high numerical aperture objective (≥ 1.2), required to visualize a single fluorophore. By axial scanning, using an electrically tunable lens with a fixed sample, we were successfully able to visualize the epidermal growth factor receptor (EGFR) moving along the three-dimensionally elongated filamentous actin bundles connecting cells (intercellular nanotube), while another EGFR on the intercellular nanotube was trapped by optical tweezers in living cells. Our approach is simple, fast and inexpensive, but it is powerful for imaging target molecules axially in single-molecule optical imaging-force spectroscopy.
DOI
Real-time optical imaging combined with single-molecule manipulation broadens the horizons for acquiring information about the spatiotemporal localization and the mechanical details of target molecules. To obtain an optical signal outside the focal plane without unintended interruption of the force signal in single-molecule optical imaging-force spectroscopy, we developed an optical method to extend the depth of field in a high numerical aperture objective (≥ 1.2), required to visualize a single fluorophore. By axial scanning, using an electrically tunable lens with a fixed sample, we were successfully able to visualize the epidermal growth factor receptor (EGFR) moving along the three-dimensionally elongated filamentous actin bundles connecting cells (intercellular nanotube), while another EGFR on the intercellular nanotube was trapped by optical tweezers in living cells. Our approach is simple, fast and inexpensive, but it is powerful for imaging target molecules axially in single-molecule optical imaging-force spectroscopy.
DOI
Testing sub-gravitational forces on atoms from a miniature in-vacuum source mass
Matt Jaffe, Philipp Haslinger, Victoria Xu, Paul Hamilton, Amol Upadhye, Benjamin Elder, Justin Khoury & Holger Müller
Traditional gravity measurements use bulk masses to both source and probe gravitational fields1. Matter-wave interferometers enable the use of probe masses as small as neutrons2, atoms3 and molecular clusters4, but still require fields generated by masses ranging from hundreds of kilograms5,6 to the entire Earth. Shrinking the sources would enable versatile configurations, improve positioning accuracy, enable tests for beyond-standard-model (‘fifth’) forces, and allow observation of non-classical effects of gravity. Here we detect the gravitational force between freely falling caesium atoms and an in-vacuum, miniature (centimetre-sized, 0.19 kg) source mass using atom interferometry. Sensitivity down to gravitational strength forces accesses the natural scale7 for a wide class of cosmologically motivated scalar field models8,9 of modified gravity and dark energy. We improve the limits on two such models, chameleons9 and symmetrons10,11, by over two orders of magnitude. We expect further tests of dark energy theories, and measurements of Newton’s gravitational constant and the gravitational Aharonov–Bohm effect12.
DOI
Traditional gravity measurements use bulk masses to both source and probe gravitational fields1. Matter-wave interferometers enable the use of probe masses as small as neutrons2, atoms3 and molecular clusters4, but still require fields generated by masses ranging from hundreds of kilograms5,6 to the entire Earth. Shrinking the sources would enable versatile configurations, improve positioning accuracy, enable tests for beyond-standard-model (‘fifth’) forces, and allow observation of non-classical effects of gravity. Here we detect the gravitational force between freely falling caesium atoms and an in-vacuum, miniature (centimetre-sized, 0.19 kg) source mass using atom interferometry. Sensitivity down to gravitational strength forces accesses the natural scale7 for a wide class of cosmologically motivated scalar field models8,9 of modified gravity and dark energy. We improve the limits on two such models, chameleons9 and symmetrons10,11, by over two orders of magnitude. We expect further tests of dark energy theories, and measurements of Newton’s gravitational constant and the gravitational Aharonov–Bohm effect12.
DOI
A minimally invasive optical trapping system to understand cellular interactions at onset of an immune response
David G. Glass, Niall McAlinden, Owain R. Millington, Amanda J. Wright
T-cells and antigen presenting cells are an essential part of the adaptive immune response system and how they interact is crucial in how the body effectively fights infection or responds to vaccines. Much of the experimental work studying interaction forces between cells has looked at the average properties of bulk samples of cells or applied microscopy to image the dynamic contact between these cells. In this paper we present a novel optical trapping technique for interrogating the force of this interaction and measuring relative interaction forces at the single-cell level. A triple-spot optical trap is used to directly manipulate the cells of interest without introducing foreign bodies such as beads to the system. The optical trap is used to directly control the initiation of cell-cell contact and, subsequently to terminate the interaction at a defined time point. The laser beam power required to separate immune cell pairs is determined and correlates with the force applied by the optical trap. As proof of concept, the antigen-specific increase in interaction force between a dendritic cell and a specific T-cell is demonstrated. Furthermore, it is demonstrated that this interaction force is completely abrogated when T-cell signalling is blocked. As a result the potential of using optical trapping to interrogate cellular interactions at the single cell level without the need to introduce foreign bodies such as beads is clearly demonstrated.
DOI
T-cells and antigen presenting cells are an essential part of the adaptive immune response system and how they interact is crucial in how the body effectively fights infection or responds to vaccines. Much of the experimental work studying interaction forces between cells has looked at the average properties of bulk samples of cells or applied microscopy to image the dynamic contact between these cells. In this paper we present a novel optical trapping technique for interrogating the force of this interaction and measuring relative interaction forces at the single-cell level. A triple-spot optical trap is used to directly manipulate the cells of interest without introducing foreign bodies such as beads to the system. The optical trap is used to directly control the initiation of cell-cell contact and, subsequently to terminate the interaction at a defined time point. The laser beam power required to separate immune cell pairs is determined and correlates with the force applied by the optical trap. As proof of concept, the antigen-specific increase in interaction force between a dendritic cell and a specific T-cell is demonstrated. Furthermore, it is demonstrated that this interaction force is completely abrogated when T-cell signalling is blocked. As a result the potential of using optical trapping to interrogate cellular interactions at the single cell level without the need to introduce foreign bodies such as beads is clearly demonstrated.
DOI
Optical trapping and optical force positioning of two-dimensional materials
M. G. Donato,E. Messina, A. Foti, T. J. Smart, P. H. Jones, M. A. Iatì, R. Saija, P. G. Gucciardi and O. M. Maragò
In recent years, considerable effort has been devoted to the synthesis and characterization of two-dimensional materials. Liquid phase exfoliation (LPE) represents a simple, large-scale method to exfoliate layered materials down to mono- and few-layer flakes. In this context, the contactless trapping, characterization, and manipulation of individual nanosheets hold perspectives for increased accuracy in flake metrology and the assembly of novel functional materials. Here, we use optical forces for high-resolution structural characterization and precise mechanical positioning of nanosheets of hexagonal boron nitride, molybdenum disulfide, and tungsten disulfide obtained by LPE. Weakly optically absorbing nanosheets of boron nitride are trapped in optical tweezers. The analysis of the thermal fluctuations allows a direct measurement of optical forces and the mean flake size in a liquid environment. Measured optical trapping constants are compared with T-matrix light scattering calculations to show a quadratic size scaling for small size, as expected for a bidimensional system. In contrast, strongly absorbing nanosheets of molybdenum disulfide and tungsten disulfide are not stably trapped due to the dominance of radiation pressure over the optical trapping force. Thus, optical forces are used to pattern a substrate by selectively depositing nanosheets in short times (minutes) and without any preparation of the surface. This study will be useful for improving ink-jet printing and for a better engineering of optoelectronic devices based on two-dimensional materials.
DOI
In recent years, considerable effort has been devoted to the synthesis and characterization of two-dimensional materials. Liquid phase exfoliation (LPE) represents a simple, large-scale method to exfoliate layered materials down to mono- and few-layer flakes. In this context, the contactless trapping, characterization, and manipulation of individual nanosheets hold perspectives for increased accuracy in flake metrology and the assembly of novel functional materials. Here, we use optical forces for high-resolution structural characterization and precise mechanical positioning of nanosheets of hexagonal boron nitride, molybdenum disulfide, and tungsten disulfide obtained by LPE. Weakly optically absorbing nanosheets of boron nitride are trapped in optical tweezers. The analysis of the thermal fluctuations allows a direct measurement of optical forces and the mean flake size in a liquid environment. Measured optical trapping constants are compared with T-matrix light scattering calculations to show a quadratic size scaling for small size, as expected for a bidimensional system. In contrast, strongly absorbing nanosheets of molybdenum disulfide and tungsten disulfide are not stably trapped due to the dominance of radiation pressure over the optical trapping force. Thus, optical forces are used to pattern a substrate by selectively depositing nanosheets in short times (minutes) and without any preparation of the surface. This study will be useful for improving ink-jet printing and for a better engineering of optoelectronic devices based on two-dimensional materials.
DOI
Attractive force on atoms due to blackbody radiation
Philipp Haslinger, Matt Jaffe, Victoria Xu, Osip Schwartz, Matthias Sonnleitner, Monika Ritsch-Marte, Helmut Ritsch & Holger Müller
Objects at finite temperature emit thermal radiation with an outward energy–momentum flow, which exerts an outward radiation pressure. At room temperature, a caesium atom scatters on average less than one of these blackbody radiation photons every 108 years. Thus, it is generally assumed that any scattering force exerted on atoms by such radiation is negligible. However, atoms also interact coherently with the thermal electromagnetic field. In this work, we measure an attractive force induced by blackbody radiation between a caesium atom and a heated, centimetre-sized cylinder, which is orders of magnitude stronger than the outward-directed radiation pressure. Using atom interferometry, we find that this force scales with the fourth power of the cylinder’s temperature. The force is in good agreement with that predicted from an a.c. Stark shift gradient of the atomic ground state in the thermal radiation field1. This observed force dominates over both gravity and radiation pressure, and does so for a large temperature range.
DOI
Objects at finite temperature emit thermal radiation with an outward energy–momentum flow, which exerts an outward radiation pressure. At room temperature, a caesium atom scatters on average less than one of these blackbody radiation photons every 108 years. Thus, it is generally assumed that any scattering force exerted on atoms by such radiation is negligible. However, atoms also interact coherently with the thermal electromagnetic field. In this work, we measure an attractive force induced by blackbody radiation between a caesium atom and a heated, centimetre-sized cylinder, which is orders of magnitude stronger than the outward-directed radiation pressure. Using atom interferometry, we find that this force scales with the fourth power of the cylinder’s temperature. The force is in good agreement with that predicted from an a.c. Stark shift gradient of the atomic ground state in the thermal radiation field1. This observed force dominates over both gravity and radiation pressure, and does so for a large temperature range.
DOI
Position-resolved Raman spectra from a laser-trapped single airborne chemical droplet
Aimable Kalume, Eric Zhu, Chuji Wang, Joshua Santarpia, and Yong-Le Pan
It could be very useful to detect and monitor the molecules and molecular reactions located at different positions within a microsized particle as they respond to various micro-local environments. In this Letter, a particular optical trap using two focusing counterpropagating hollow beams was able to stably trap both absorbing and nonabsorbing particles in air for lengthy observation. A technique that can measure the Raman spectra from different submicrometer positions of a laser-trapped single airborne particle was developed. Spontaneous and stimulated Raman scattering spectra originating from different positions of a diethyl phthalate droplet were recorded, and the strong Raman scattering signals are the result of cavity-enhanced effects and the localized strong light illumination.
DOI
It could be very useful to detect and monitor the molecules and molecular reactions located at different positions within a microsized particle as they respond to various micro-local environments. In this Letter, a particular optical trap using two focusing counterpropagating hollow beams was able to stably trap both absorbing and nonabsorbing particles in air for lengthy observation. A technique that can measure the Raman spectra from different submicrometer positions of a laser-trapped single airborne particle was developed. Spontaneous and stimulated Raman scattering spectra originating from different positions of a diethyl phthalate droplet were recorded, and the strong Raman scattering signals are the result of cavity-enhanced effects and the localized strong light illumination.
DOI
Tuesday, January 16, 2018
Impact of complex surfaces on biomicrorheological measurements using optical tweezers
Shu Zhang, Lachlan J. Gibson, Alexander B. Stilgoe, Timo A. Nieminen and Halina Rubinsztein-Dunlop
The characterisation of physical properties in biologically relevant processes and the development of novel microfluidic devices for this purpose are experiencing a great resurgence at present. In many of measurements of this type where a probe in a fluid is used, the strong influence of the boundaries of the volume used is a serious problem. In these geometries the proximity of a probe to a wall can severely influence the measurement. However, although much knowledge has been gained about flat walls, to date, the effect of non-planar surfaces at microscopic scale on rotational motion of micro-objects has not been studied. Here we present for the first time both experimental measurements and numerical computations which aim to study the drag torque on optically trapped rotating particles moving near 3D-printed conical and cylindrical walls on-chip. These results are essential for quantifying how curved walls can effect the torque on particles, and thus enable accurate hydrodynamic simulations at the micron-scale. This opens the potential for new sensing approaches under more complex conditions, allowing both dynamic and microrheological studies of biological systems and lab-on-chip devices.
DOI
The characterisation of physical properties in biologically relevant processes and the development of novel microfluidic devices for this purpose are experiencing a great resurgence at present. In many of measurements of this type where a probe in a fluid is used, the strong influence of the boundaries of the volume used is a serious problem. In these geometries the proximity of a probe to a wall can severely influence the measurement. However, although much knowledge has been gained about flat walls, to date, the effect of non-planar surfaces at microscopic scale on rotational motion of micro-objects has not been studied. Here we present for the first time both experimental measurements and numerical computations which aim to study the drag torque on optically trapped rotating particles moving near 3D-printed conical and cylindrical walls on-chip. These results are essential for quantifying how curved walls can effect the torque on particles, and thus enable accurate hydrodynamic simulations at the micron-scale. This opens the potential for new sensing approaches under more complex conditions, allowing both dynamic and microrheological studies of biological systems and lab-on-chip devices.
DOI
Three-dimensional holographic optical manipulation through a high-numerical-aperture soft-glass multimode fibre
Ivo T. Leite, Sergey Turtaev, Xin Jiang, Martin Šiler, Alfred Cuschieri, Philip St. J. Russell & Tomáš Čižmár
Holographic optical tweezers (HOT) hold great promise for many applications in biophotonics, allowing the creation and measurement of minuscule forces on biomolecules, molecular motors and cells. Geometries used in HOT currently rely on bulk optics, and their exploitation in vivo is compromised by the optically turbid nature of tissues. We present an alternative HOT approach in which multiple three-dimensional (3D) traps are introduced through a high-numerical-aperture multimode optical fibre, thus enabling an equally versatile means of manipulation through channels having cross-section comparable to the size of a single cell. Our work demonstrates real-time manipulation of 3D arrangements of micro-objects, as well as manipulation inside otherwise inaccessible cavities. We show that the traps can be formed over fibre lengths exceeding 100 mm and positioned with nanometric resolution. The results provide the basis for holographic manipulation and other high-numerical-aperture techniques, including advanced microscopy, through single-core-fibre endoscopes deep inside living tissues and other complex environments.
DOI
Holographic optical tweezers (HOT) hold great promise for many applications in biophotonics, allowing the creation and measurement of minuscule forces on biomolecules, molecular motors and cells. Geometries used in HOT currently rely on bulk optics, and their exploitation in vivo is compromised by the optically turbid nature of tissues. We present an alternative HOT approach in which multiple three-dimensional (3D) traps are introduced through a high-numerical-aperture multimode optical fibre, thus enabling an equally versatile means of manipulation through channels having cross-section comparable to the size of a single cell. Our work demonstrates real-time manipulation of 3D arrangements of micro-objects, as well as manipulation inside otherwise inaccessible cavities. We show that the traps can be formed over fibre lengths exceeding 100 mm and positioned with nanometric resolution. The results provide the basis for holographic manipulation and other high-numerical-aperture techniques, including advanced microscopy, through single-core-fibre endoscopes deep inside living tissues and other complex environments.
DOI
“Freezing” of NaClO3 Metastable Crystalline State by Optical Trapping in Unsaturated Microdroplet
Hiromasa Niinomi, Teruki Sugiyama, Katsuhiko Miyamoto, and Takashige Omatsu
We reversibly controlled phase conversion between a microdroplet of a NaClO3 unsaturated aqueous solution and a metastable single crystal, which is usually a short-lived phase in spontaneous crystallization, simply by irradiating a tightly focused visible continuous-wave (CW) laser to the microdroplet. The laser irradiation allowed the metastable crystal to generate and stably grow without a polymorphic transformation. This successful metastable phase control is attributed to the combination of the advantage of optical trapping-induced nucleation that nucleation takes place from unsaturated mother solution and the advantage of microdroplet method, which suppresses additional nucleation leading to the transformation. In situ observation shows the crystal dissolves when the laser irradiation is stopped, whereas the laser irradiation stabilizes the crystal even if the size of the crystal becomes larger than that of focal spot. These observations indicate that a change in the relative magnitudes of chemical potentials between solution/crystalline phases. This change is possibly promoted via crystal growth by trapping of crystalline clusters in optical potential well formed on a crystal surfaces originating from “light propagation” through the crystal. Our results shed a light not only on polymorph control but also on a method to prepare a longer-lived achiral precursor for analysis on achiral–chiral transition by “freezing” a kinetic pathway of chiral crystallization.
DOI
We reversibly controlled phase conversion between a microdroplet of a NaClO3 unsaturated aqueous solution and a metastable single crystal, which is usually a short-lived phase in spontaneous crystallization, simply by irradiating a tightly focused visible continuous-wave (CW) laser to the microdroplet. The laser irradiation allowed the metastable crystal to generate and stably grow without a polymorphic transformation. This successful metastable phase control is attributed to the combination of the advantage of optical trapping-induced nucleation that nucleation takes place from unsaturated mother solution and the advantage of microdroplet method, which suppresses additional nucleation leading to the transformation. In situ observation shows the crystal dissolves when the laser irradiation is stopped, whereas the laser irradiation stabilizes the crystal even if the size of the crystal becomes larger than that of focal spot. These observations indicate that a change in the relative magnitudes of chemical potentials between solution/crystalline phases. This change is possibly promoted via crystal growth by trapping of crystalline clusters in optical potential well formed on a crystal surfaces originating from “light propagation” through the crystal. Our results shed a light not only on polymorph control but also on a method to prepare a longer-lived achiral precursor for analysis on achiral–chiral transition by “freezing” a kinetic pathway of chiral crystallization.
DOI
Stretching and Heating Single DNA Molecules with Optically Trapped Gold–Silica Janus Particles
Sabrina Simoncelli, Samuel Johnson, Franziska Kriegel, Jan Lipfert, and Jochen Feldmann
Self-propelled micro- and nanoscale motors are capable of autonomous motion typically by inducing local concentration gradients or thermal gradients in their surrounding medium. This is a result of the heterogeneous surface of the self-propelled structures that consist of materials with different chemical or physical properties. Here we present a self-thermophoretically driven Au–silica Janus particle that can simultaneously stretch and partially melt a single double-stranded DNA molecule. We show that the effective force acting on the DNA molecule is in the ∼pN range, well suited to probe the entropic stretching regime of DNA, and we demonstrate that the local temperature enhancement around the gold side of the particle produces partial DNA dehybridization.
DOI
Self-propelled micro- and nanoscale motors are capable of autonomous motion typically by inducing local concentration gradients or thermal gradients in their surrounding medium. This is a result of the heterogeneous surface of the self-propelled structures that consist of materials with different chemical or physical properties. Here we present a self-thermophoretically driven Au–silica Janus particle that can simultaneously stretch and partially melt a single double-stranded DNA molecule. We show that the effective force acting on the DNA molecule is in the ∼pN range, well suited to probe the entropic stretching regime of DNA, and we demonstrate that the local temperature enhancement around the gold side of the particle produces partial DNA dehybridization.
DOI
The energy cost of polypeptide knot formation and its folding consequences
Andrés Bustamante, Juan Sotelo-Campos, Daniel G. Guerra, Martin Floor, Christian A. M. Wilson, Carlos Bustamante & Mauricio Báez
Knots are natural topologies of chains. Yet, little is known about spontaneous knot formation in a polypeptide chain—an event that can potentially impair its folding—and about the effect of a knot on the stability and folding kinetics of a protein. Here we used optical tweezers to show that the free energy cost to form a trefoil knot in the denatured state of a polypeptide chain of 120 residues is 5.8 ± 1 kcal mol−1. Monte Carlo dynamics of random chains predict this value, indicating that the free energy cost of knot formation is of entropic origin. This cost is predicted to remain above 3 kcal mol−1 for denatured proteins as large as 900 residues. Therefore, we conclude that naturally knotted proteins cannot attain their knot randomly in the unfolded state but must pay the cost of knotting through contacts along their folding landscape.
DOI
Knots are natural topologies of chains. Yet, little is known about spontaneous knot formation in a polypeptide chain—an event that can potentially impair its folding—and about the effect of a knot on the stability and folding kinetics of a protein. Here we used optical tweezers to show that the free energy cost to form a trefoil knot in the denatured state of a polypeptide chain of 120 residues is 5.8 ± 1 kcal mol−1. Monte Carlo dynamics of random chains predict this value, indicating that the free energy cost of knot formation is of entropic origin. This cost is predicted to remain above 3 kcal mol−1 for denatured proteins as large as 900 residues. Therefore, we conclude that naturally knotted proteins cannot attain their knot randomly in the unfolded state but must pay the cost of knotting through contacts along their folding landscape.
DOI
Optical Forces at the Nanoscale: Size and Electrostatic Effects
Paloma Rodríguez-Sevilla, Katarzyna Prorok, Artur Bednarkiewicz, Manuel I. Marqués, Antonio García-Martín, José García Solé, Patricia Haro-González, and Daniel Jaque
The reduced magnitude of the optical trapping forces exerted over sub-200 nm dielectric nanoparticles complicates their optical manipulation, hindering the development of techniques and studies based on it. Improvement of trapping capabilities for such tiny objects requires a deep understanding of the mechanisms beneath them. Traditionally, the optical forces acting on dielectric nanoparticles have been only correlated with their volume, and the size has been traditionally identified as a key parameter. However, the most recently published research results have shown that the electrostatic characteristics of a sub-100 nm dielectric particle could also play a significant role. Indeed, at present it is not clear what optical forces depend. In this work, we designed a set of experiments in order to elucidate the different mechanism and properties (i.e., size and/or electrostatic properties) that governs the magnitude of optical forces. The comparison between experimental data and numerical simulations have shown that the double layer induced at nanoparticle’s surface, not considered in the classical description of nanoparticle’s polarizability, plays a relevant role determining the magnitude of the optical forces. Here, the presented results constitute the first step toward the development of the dielectric nanoparticle over which enhanced optical forces could be exerted, enabling their optical manipulation for multiples purposes ranging from fundamental to applied studies.
DOI
The reduced magnitude of the optical trapping forces exerted over sub-200 nm dielectric nanoparticles complicates their optical manipulation, hindering the development of techniques and studies based on it. Improvement of trapping capabilities for such tiny objects requires a deep understanding of the mechanisms beneath them. Traditionally, the optical forces acting on dielectric nanoparticles have been only correlated with their volume, and the size has been traditionally identified as a key parameter. However, the most recently published research results have shown that the electrostatic characteristics of a sub-100 nm dielectric particle could also play a significant role. Indeed, at present it is not clear what optical forces depend. In this work, we designed a set of experiments in order to elucidate the different mechanism and properties (i.e., size and/or electrostatic properties) that governs the magnitude of optical forces. The comparison between experimental data and numerical simulations have shown that the double layer induced at nanoparticle’s surface, not considered in the classical description of nanoparticle’s polarizability, plays a relevant role determining the magnitude of the optical forces. Here, the presented results constitute the first step toward the development of the dielectric nanoparticle over which enhanced optical forces could be exerted, enabling their optical manipulation for multiples purposes ranging from fundamental to applied studies.
DOI
Methods of Micropatterning and Manipulation of Cells for Biomedical Applications
Adrian Martinez-Rivas, Génesis K. González-Quijano, Sergio Proa-Coronado, Childérick Séverac and Etienne Dague
Micropatterning and manipulation of mammalian and bacterial cells are important in biomedical studies to perform in vitro assays and to evaluate biochemical processes accurately, establishing the basis for implementing biomedical microelectromechanical systems (bioMEMS), point-of-care (POC) devices, or organs-on-chips (OOC), which impact on neurological, oncological, dermatologic, or tissue engineering issues as part of personalized medicine. Cell patterning represents a crucial step in fundamental and applied biological studies in vitro, hence today there are a myriad of materials and techniques that allow one to immobilize and manipulate cells, imitating the 3D in vivo milieu. This review focuses on current physical cell patterning, plus chemical and a combination of them both that utilizes different materials and cutting-edge micro-nanofabrication methodologies.
DOI
Micropatterning and manipulation of mammalian and bacterial cells are important in biomedical studies to perform in vitro assays and to evaluate biochemical processes accurately, establishing the basis for implementing biomedical microelectromechanical systems (bioMEMS), point-of-care (POC) devices, or organs-on-chips (OOC), which impact on neurological, oncological, dermatologic, or tissue engineering issues as part of personalized medicine. Cell patterning represents a crucial step in fundamental and applied biological studies in vitro, hence today there are a myriad of materials and techniques that allow one to immobilize and manipulate cells, imitating the 3D in vivo milieu. This review focuses on current physical cell patterning, plus chemical and a combination of them both that utilizes different materials and cutting-edge micro-nanofabrication methodologies.
DOI
Monday, January 15, 2018
Light-driven self-assembly of hetero-shaped gold nanorods
Jiunn-Woei Liaw, Hsueh-Yu Chao, Cheng-Wei Huang, Mao-Kuen Kuo
Light-driven self-assembly and coalescence of two nearby hetero-shaped gold nanorods (GNRs) with different lengths are studied theoretically. The optical forces and torques, in terms of Maxwell’s stress tensor, upon these GNRs provided by a linearly polarized (LP) plane wave are analyzed using the multiple multipole (MMP) method. Numerical results show that the optical torque dominates their alignments and the optical force their attraction. The most likely outcome of the plasmon-mediated light–matter interaction is wavelength dependent. Three different coalescences of the two GNRs could be induced by a LP light in three different wavelength regimes, respectively. For example, the side-by-side coalescence of two GNRs with radius of 15 nm and different lengths (120 and 240 nm) is induced in water as irradiated by a LP light at 633 nm, the T-shaped one at 1064 nm, and the end-to-end one at 1700 nm. The plasmonic attractive force and heating power densities inside GNRs with different gaps are also studied; the smaller the gap, the larger the attractive force and heating power. The results imply that the plasmonic coalescence and heating of two discrete GNRs may cause the local fusion at the junction of the assembly and the subsequent annealing (even recrystallization). Because the heating makes the two discrete GNRs fused to become a new nanostructure, the plasmonic coalescence of optical manipulation is irreversible.
DOI
Light-driven self-assembly and coalescence of two nearby hetero-shaped gold nanorods (GNRs) with different lengths are studied theoretically. The optical forces and torques, in terms of Maxwell’s stress tensor, upon these GNRs provided by a linearly polarized (LP) plane wave are analyzed using the multiple multipole (MMP) method. Numerical results show that the optical torque dominates their alignments and the optical force their attraction. The most likely outcome of the plasmon-mediated light–matter interaction is wavelength dependent. Three different coalescences of the two GNRs could be induced by a LP light in three different wavelength regimes, respectively. For example, the side-by-side coalescence of two GNRs with radius of 15 nm and different lengths (120 and 240 nm) is induced in water as irradiated by a LP light at 633 nm, the T-shaped one at 1064 nm, and the end-to-end one at 1700 nm. The plasmonic attractive force and heating power densities inside GNRs with different gaps are also studied; the smaller the gap, the larger the attractive force and heating power. The results imply that the plasmonic coalescence and heating of two discrete GNRs may cause the local fusion at the junction of the assembly and the subsequent annealing (even recrystallization). Because the heating makes the two discrete GNRs fused to become a new nanostructure, the plasmonic coalescence of optical manipulation is irreversible.
DOI
Modified field confinement and enhanced optical forces in hybrid dielectric wedge tip-loaded plasmonic waveguide
Xiaogang Chen; Qijing Lu; Hongqin Yang; Xiang Wu; Shusen Xie;
We proposed and theoretically investigated a hybrid plasmonics waveguide consisting of a tiptilted quadrate nanowire, which was embedded in a low-permittivity dielectric and placed on a metal substrate with a small gap distance. Due to the corner effect, the hybrid mode with extremely local field enhancement has the long propagation length and strong coupling strength between the dielectric nanowire and metal. By employing the simulations with different geometric parameters, the proposed waveguide can obtain better performances than the previous hybrid plasmonics waveguide, particularly in the subwavelength confinement (as small as λ^2/1600 ), long-range propagation (millimeter range), and optical trapping forces ( 2.12 pN/W). The use of a naturally dielectric wedge tip of quadrate nanowire that can be chemically synthesized provides an efficient approach to circumvent the fabrication difficulty of shape wedge tips. The present structure provides an excellent platform for nanophotonic waveguides, nanolasers, nanoscale optical tweezers, and biosensing.
DOI
We proposed and theoretically investigated a hybrid plasmonics waveguide consisting of a tiptilted quadrate nanowire, which was embedded in a low-permittivity dielectric and placed on a metal substrate with a small gap distance. Due to the corner effect, the hybrid mode with extremely local field enhancement has the long propagation length and strong coupling strength between the dielectric nanowire and metal. By employing the simulations with different geometric parameters, the proposed waveguide can obtain better performances than the previous hybrid plasmonics waveguide, particularly in the subwavelength confinement (as small as λ^2/1600 ), long-range propagation (millimeter range), and optical trapping forces ( 2.12 pN/W). The use of a naturally dielectric wedge tip of quadrate nanowire that can be chemically synthesized provides an efficient approach to circumvent the fabrication difficulty of shape wedge tips. The present structure provides an excellent platform for nanophotonic waveguides, nanolasers, nanoscale optical tweezers, and biosensing.
DOI
Effects of finite and discrete sampling and blur on microrheology experiments
Victoria E. Loosemore and Nancy R. Forde
The frequency-dependent viscous and elastic properties of fluids can be determined from measurements of the thermal fluctuations of a micron-sized particle trapped by optical tweezers. Finite bandwidth and other instrument limitations lead to systematic errors in measurement of the fluctuations. In this work, we numerically represented power spectra of bead position measurements as if collected by two different measurement devices: a quadrant photodiode, which measures the deflection of the trapping laser; and a high-speed camera, which images the trapped bead directly. We explored the effects of aliasing, camera blur, sampling frequency, and measurement time. By comparing the power spectrum, complex response function, and the complex shear modulus with the ideal values, we found that the viscous and elastic properties inferred from the data are affected by the instrument limitations of each device. We discuss how these systematic effects might affect experimental results from microrheology measurements and suggest approaches to reduce discrepancies.
DOI
The frequency-dependent viscous and elastic properties of fluids can be determined from measurements of the thermal fluctuations of a micron-sized particle trapped by optical tweezers. Finite bandwidth and other instrument limitations lead to systematic errors in measurement of the fluctuations. In this work, we numerically represented power spectra of bead position measurements as if collected by two different measurement devices: a quadrant photodiode, which measures the deflection of the trapping laser; and a high-speed camera, which images the trapped bead directly. We explored the effects of aliasing, camera blur, sampling frequency, and measurement time. By comparing the power spectrum, complex response function, and the complex shear modulus with the ideal values, we found that the viscous and elastic properties inferred from the data are affected by the instrument limitations of each device. We discuss how these systematic effects might affect experimental results from microrheology measurements and suggest approaches to reduce discrepancies.
DOI
Partially native intermediates mediate misfolding of SOD1 in single-molecule folding trajectories
Supratik Sen Mojumdar, Zackary N. Scholl, Derek R. Dee, Logan Rouleau, Uttam Anand, Craig Garen & Michael T. Woodside
Prion-like misfolding of superoxide dismutase 1 (SOD1) is associated with the disease ALS, but the mechanism of misfolding remains unclear, partly because misfolding is difficult to observe directly. Here we study the most misfolding-prone form of SOD1, reduced un-metallated monomers, using optical tweezers to measure unfolding and refolding of single molecules. We find that the folding is more complex than suspected, resolving numerous previously undetected intermediate states consistent with the formation of individual β-strands in the native structure. We identify a stable core of the protein that unfolds last and refolds first, and directly observe several distinct misfolded states that branch off from the native folding pathways at specific points after the formation of the stable core. Partially folded intermediates thus play a crucial role mediating between native and non-native folding. These results suggest an explanation for SOD1’s propensity for prion-like misfolding and point to possible targets for therapeutic intervention.
DOI
Prion-like misfolding of superoxide dismutase 1 (SOD1) is associated with the disease ALS, but the mechanism of misfolding remains unclear, partly because misfolding is difficult to observe directly. Here we study the most misfolding-prone form of SOD1, reduced un-metallated monomers, using optical tweezers to measure unfolding and refolding of single molecules. We find that the folding is more complex than suspected, resolving numerous previously undetected intermediate states consistent with the formation of individual β-strands in the native structure. We identify a stable core of the protein that unfolds last and refolds first, and directly observe several distinct misfolded states that branch off from the native folding pathways at specific points after the formation of the stable core. Partially folded intermediates thus play a crucial role mediating between native and non-native folding. These results suggest an explanation for SOD1’s propensity for prion-like misfolding and point to possible targets for therapeutic intervention.
DOI
The Role of Nanomechanics in Healthcare
Pranjal Nautiyal, Fahad Alam, Kantesh Balani, Arvind Agarwal
Nanomechanics has played a vital role in pushing our capability to detect, probe, and manipulate the biological species, such as proteins, cells, and tissues, paving way to a deeper knowledge and superior strategies for healthcare. Nanomechanical characterization techniques, such as atomic force microscopy, nanoindentation, nanotribology, optical tweezers, and other hybrid techniques have been utilized to understand the mechanics and kinetics of biospecies. Investigation of the mechanics of cells and tissues has provided critical information about mechanical characteristics of host body environments. This information has been utilized for developing biomimetic materials and structures for tissue engineering and artificial implants. This review summarizes nanomechanical characterization techniques and their potential applications in healthcare research. The principles and examples of label-free detection of cancers and myocardial infarction by nanomechanical cantilevers are discussed. The vital importance of nanomechanics in regenerative medicine is highlighted from the perspective of material selection and design for developing biocompatible scaffolds. This review interconnects the advancements made in fundamental materials science research and biomedical technology, and therefore provides scientific insight that is of common interest to the researchers working in different disciplines of healthcare science and technology.
DOI
Nanomechanics has played a vital role in pushing our capability to detect, probe, and manipulate the biological species, such as proteins, cells, and tissues, paving way to a deeper knowledge and superior strategies for healthcare. Nanomechanical characterization techniques, such as atomic force microscopy, nanoindentation, nanotribology, optical tweezers, and other hybrid techniques have been utilized to understand the mechanics and kinetics of biospecies. Investigation of the mechanics of cells and tissues has provided critical information about mechanical characteristics of host body environments. This information has been utilized for developing biomimetic materials and structures for tissue engineering and artificial implants. This review summarizes nanomechanical characterization techniques and their potential applications in healthcare research. The principles and examples of label-free detection of cancers and myocardial infarction by nanomechanical cantilevers are discussed. The vital importance of nanomechanics in regenerative medicine is highlighted from the perspective of material selection and design for developing biocompatible scaffolds. This review interconnects the advancements made in fundamental materials science research and biomedical technology, and therefore provides scientific insight that is of common interest to the researchers working in different disciplines of healthcare science and technology.
DOI
Chiral liquid crystal colloids
Ye Yuan, Angel Martinez, Bohdan Senyuk, Mykola Tasinkevych & Ivan I. Smalyukh
Colloidal particles disturb the alignment of rod-like molecules of liquid crystals, giving rise to long-range interactions that minimize the free energy of distorted regions. Particle shape and topology are known to guide this self-assembly process. However, how chirality of colloidal inclusions affects these long-range interactions is unclear. Here we study the effects of distortions caused by chiral springs and helices on the colloidal self-organization in a nematic liquid crystal using laser tweezers, particle tracking and optical imaging. We show that chirality of colloidal particles interacts with the nematic elasticity to predefine chiral or racemic colloidal superstructures in nematic colloids. These findings are consistent with numerical modelling based on the minimization of Landau–de Gennes free energy. Our study uncovers the role of chirality in defining the mesoscopic order of liquid crystal colloids, suggesting that this feature may be a potential tool to modulate the global orientated self-organization of these systems.
DOI
Colloidal particles disturb the alignment of rod-like molecules of liquid crystals, giving rise to long-range interactions that minimize the free energy of distorted regions. Particle shape and topology are known to guide this self-assembly process. However, how chirality of colloidal inclusions affects these long-range interactions is unclear. Here we study the effects of distortions caused by chiral springs and helices on the colloidal self-organization in a nematic liquid crystal using laser tweezers, particle tracking and optical imaging. We show that chirality of colloidal particles interacts with the nematic elasticity to predefine chiral or racemic colloidal superstructures in nematic colloids. These findings are consistent with numerical modelling based on the minimization of Landau–de Gennes free energy. Our study uncovers the role of chirality in defining the mesoscopic order of liquid crystal colloids, suggesting that this feature may be a potential tool to modulate the global orientated self-organization of these systems.
DOI
Lateral sorting of chiral nanoparticles using Fano-enhanced chiral force in visible region
Tun Cao and Yimei Qiu
Chiral gradient force allows a passive separation of an enantiomer since its direction is dependent on the handedness of its chiral entities. However, chiral polarisability is much weaker than electric polarisability. As a consequence, the non-chiral gradient force dominates over chiral force, which makes enantioselective sorting challenging. We present here, both numerically and analytically, that the chiral gradient force acting on chiral nanoparticles can overcome the non-chiral force when specimens are placed in a Fano-enhanced chiral gradient near-field using a plasmonic nanoaperture. Under circularly polarized light illumination, the interaction between the resonant modes of symmetric outer and asymmetric inner Au split-rings results in a splitting of the modal energies, which excites multipolar interference Fano resonances (FRs). This enables a local aperture between the two split-rings to possess very large optical chirality gradients while maintaining low gradients of electromagnetic energy density around the FRs from the visible region. By way of the lateral resultant force composed of both chiral and non-chiral gradient forces, we can accomplish a helicity-dependent transverse deflection of the chiral nanoparticles positioned above the aperture, which may offer a good platform for all-optical enantiopure compounds.
DOI
Chiral gradient force allows a passive separation of an enantiomer since its direction is dependent on the handedness of its chiral entities. However, chiral polarisability is much weaker than electric polarisability. As a consequence, the non-chiral gradient force dominates over chiral force, which makes enantioselective sorting challenging. We present here, both numerically and analytically, that the chiral gradient force acting on chiral nanoparticles can overcome the non-chiral force when specimens are placed in a Fano-enhanced chiral gradient near-field using a plasmonic nanoaperture. Under circularly polarized light illumination, the interaction between the resonant modes of symmetric outer and asymmetric inner Au split-rings results in a splitting of the modal energies, which excites multipolar interference Fano resonances (FRs). This enables a local aperture between the two split-rings to possess very large optical chirality gradients while maintaining low gradients of electromagnetic energy density around the FRs from the visible region. By way of the lateral resultant force composed of both chiral and non-chiral gradient forces, we can accomplish a helicity-dependent transverse deflection of the chiral nanoparticles positioned above the aperture, which may offer a good platform for all-optical enantiopure compounds.
DOI
Subscribe to:
Posts (Atom)