Nima Khatibzadeh, Alexander B. Stilgoe, Ann A. M. Bui, Yesenia Rocha, Gladys M. Cruz, Vince Loke, Linda Z. Shi, Timo A. Nieminen, Halina Rubinsztein-Dunlop & Michael W. Berns
Quantitative determination of the motility forces of chromosomes during cell division is fundamental to understanding a process that is universal among eukaryotic organisms. Using an optical tweezers system, isolated mammalian chromosomes were held in a 1064 nm laser trap. The minimum force required to move a single chromosome was determined to be ≈0.8–5 pN. The maximum transverse trapping efficiency of the isolated chromosomes was calculated as ≈0.01–0.02. These results confirm theoretical force calculations of ≈0.1–12 pN to move a chromosome on the mitotic or meiotic spindle. The verification of these results was carried out by calibration of the optical tweezers when trapping microspheres with a diameter of 4.5–15 µm in media with 1–7 cP viscosity. The results of the chromosome and microsphere trapping experiments agree with optical models developed to simulate trapping of cylindrical and spherical specimens.
DOI
Thursday, November 6, 2014
The minimal cadherin-catenin complex binds to actin filaments under force
Craig D. Buckley, Jiongyi Tan, Karen L. Anderson, Dorit Hanein, Niels Volkmann, William I. Weis, W. James Nelson, Alexander R. Dunn
Cadherins are an ancient class of transmembrane proteins that are essential for the formation of multicellular tissues in metazoans. Cadherins link intercellular adhesions to the cellular cytoskeleton, but how they are connected specifically to actin filaments is a hotly debated issue. Genetic and cell culture experiments indicate that E-cadherin, β-catenin, and the actin filament binding protein αE-catenin form a minimal cadherin-catenin complex that binds to the actin cytoskeleton directly in epithelial tissues. However, experiments with purified proteins showed that a stable cadherin-catenin complex can be reconstituted, but it does not bind strongly to actin filaments in solution. Nevertheless, cell culture experiments indicated that the cadherin-catenin complex is under constitutive actomyosin-generated tension and that this connection is required for mechanotransduction at cadherin-based adhesions. Here, we tested the hypothesis that tension is required to stabilize a linkage between the cadherin-catenin complex and actin filaments, and clarify how the cadherin-catenin complex could interact directly with the actin cytoskeleton in cells.
DOI
Cadherins are an ancient class of transmembrane proteins that are essential for the formation of multicellular tissues in metazoans. Cadherins link intercellular adhesions to the cellular cytoskeleton, but how they are connected specifically to actin filaments is a hotly debated issue. Genetic and cell culture experiments indicate that E-cadherin, β-catenin, and the actin filament binding protein αE-catenin form a minimal cadherin-catenin complex that binds to the actin cytoskeleton directly in epithelial tissues. However, experiments with purified proteins showed that a stable cadherin-catenin complex can be reconstituted, but it does not bind strongly to actin filaments in solution. Nevertheless, cell culture experiments indicated that the cadherin-catenin complex is under constitutive actomyosin-generated tension and that this connection is required for mechanotransduction at cadherin-based adhesions. Here, we tested the hypothesis that tension is required to stabilize a linkage between the cadherin-catenin complex and actin filaments, and clarify how the cadherin-catenin complex could interact directly with the actin cytoskeleton in cells.
DOI
Optical trapping in the presence of higher order mode sources and interactions
I Liberal, I Ederra, R Gonzalo and R W Ziolkowski
The force fields produced by higher order mode (HOM) TMen1 sources are investigated analytically and numerically. Their application to optical manipulation and trapping is emphasized. It is found that the strong reactive fields excited by these advanced HOM nanoantennas significantly enhance both the gradient and near-field absorption-scattering forces, as well as increase the range of attraction of those absorption-scattering forces. Moreover, the impact on these force fields by the excitation of HOMs in a nanoparticle is addressed. These results are used to demonstrate the theoretical possibility of the generation of tractor beams by highly subwavelength sources on resonant dipolar nanoparticles. The analysis reveals that the associated attractive forces can be enhanced by several orders of magnitude before the HOMs induce any noticeable degradation of the radiation efficiency. The practical implementation of HOM TMen1 sources by means of active core-shell resonators, as well as their potential for the self-induced trapping of fluorescent molecules, is also addressed.
DOI
The force fields produced by higher order mode (HOM) TMen1 sources are investigated analytically and numerically. Their application to optical manipulation and trapping is emphasized. It is found that the strong reactive fields excited by these advanced HOM nanoantennas significantly enhance both the gradient and near-field absorption-scattering forces, as well as increase the range of attraction of those absorption-scattering forces. Moreover, the impact on these force fields by the excitation of HOMs in a nanoparticle is addressed. These results are used to demonstrate the theoretical possibility of the generation of tractor beams by highly subwavelength sources on resonant dipolar nanoparticles. The analysis reveals that the associated attractive forces can be enhanced by several orders of magnitude before the HOMs induce any noticeable degradation of the radiation efficiency. The practical implementation of HOM TMen1 sources by means of active core-shell resonators, as well as their potential for the self-induced trapping of fluorescent molecules, is also addressed.
DOI
Tuesday, November 4, 2014
All-fiber self-accelerating Bessel-like beam generator and its application
Zhihai Liu, Yaxun Zhang, Yu Zhang, Peibo Liang, Jun Yang, and Libo Yuan
We demonstrate an all-fiber transverse self-accelerating Bessel-like beam generator and its optical trapping application. The theoretical and experimental studies have been provided to verify this beam properties. We produce the Bessel-like beam by splicing the single-mode fiber and multimode fiber with a defined offset and then modulating the output light beam phase by fabricating a small hemispherical-lens fiber tip; therefore, the phase-modulated Bessel-like beam generates the properties of transverse self-accelerating. The transverse acceleration of the the Bessel-like beam generated here is ∼10−4 μm−1, which is almost 100 times larger than that of the beam generated in the free-space optical circuit based on the lens. The experimental and simulated results have good consistencies. The realization of the microparticle transverse acceleration transporting with this Bessel-like beam provides a new method for microparticles to be transported in a bending trajectory. This all-fiber transverse self-accelerating Bessel-like beam generator structure is simple, with high integration and small size, and constitutes a new development for high-precision biological cell experiments and manipulations.
DOI
We demonstrate an all-fiber transverse self-accelerating Bessel-like beam generator and its optical trapping application. The theoretical and experimental studies have been provided to verify this beam properties. We produce the Bessel-like beam by splicing the single-mode fiber and multimode fiber with a defined offset and then modulating the output light beam phase by fabricating a small hemispherical-lens fiber tip; therefore, the phase-modulated Bessel-like beam generates the properties of transverse self-accelerating. The transverse acceleration of the the Bessel-like beam generated here is ∼10−4 μm−1, which is almost 100 times larger than that of the beam generated in the free-space optical circuit based on the lens. The experimental and simulated results have good consistencies. The realization of the microparticle transverse acceleration transporting with this Bessel-like beam provides a new method for microparticles to be transported in a bending trajectory. This all-fiber transverse self-accelerating Bessel-like beam generator structure is simple, with high integration and small size, and constitutes a new development for high-precision biological cell experiments and manipulations.
DOI
Optical trapping reveals propulsion forces, power generation and motility efficiency of the unicellular parasites Trypanosoma brucei brucei
Eric Stellamanns, Sravanti Uppaluri, Axel Hochstetter, Niko Heddergott, Markus Engstler & Thomas Pfohl
Unicellular parasites have developed sophisticated swimming mechanisms to survive in a wide range of environments. Cell motility of African trypanosomes, parasites responsible for fatal illness in humans and animals, is crucial both in the insect vector and the mammalian host. Using millisecond-scale imaging in a microfluidics platform along with a custom made optical trap, we are able to confine single cells to study trypanosome motility. From the trapping characteristics of the cells, we determine the propulsion force generated by cells with a single flagellum as well as of dividing trypanosomes with two fully developed flagella. Estimates of the dissipative energy and the power generation of single cells obtained from the motility patterns of the trypanosomes within the optical trap indicate that specific motility characteristics, in addition to locomotion, may be required for antibody clearance. Introducing a steerable second optical trap we could further measure the force, which is generated at the flagellar tip. Differences in the cellular structure of the trypanosomes are correlated with the trapping and motility characteristics and in consequence with their propulsion force, dissipative energy and power generation.
DOI
Unicellular parasites have developed sophisticated swimming mechanisms to survive in a wide range of environments. Cell motility of African trypanosomes, parasites responsible for fatal illness in humans and animals, is crucial both in the insect vector and the mammalian host. Using millisecond-scale imaging in a microfluidics platform along with a custom made optical trap, we are able to confine single cells to study trypanosome motility. From the trapping characteristics of the cells, we determine the propulsion force generated by cells with a single flagellum as well as of dividing trypanosomes with two fully developed flagella. Estimates of the dissipative energy and the power generation of single cells obtained from the motility patterns of the trypanosomes within the optical trap indicate that specific motility characteristics, in addition to locomotion, may be required for antibody clearance. Introducing a steerable second optical trap we could further measure the force, which is generated at the flagellar tip. Differences in the cellular structure of the trypanosomes are correlated with the trapping and motility characteristics and in consequence with their propulsion force, dissipative energy and power generation.
DOI
Anomalous optical forces on the anisotropic Rayleigh particles
Y. X. Ni, J. K. Chen, and L. Gao
We investigate the optical forces on the radially anisotropic spheres from an incident plane wave based on our generalized full-wave scattering theory and the Maxwell stress tensor integration techniques. We demonstrate that the optical force on the Rayleigh sphere with radial anisotropy does not always obey the well-known Rayleigh’s law F~k40a6 (where k0 is the wave number and a is the radius of the sphere), but could be anomalous with the laws such as F~k00a2, F~k−20a0, and F~k80a10 under certain conditions. Therefore, the optical force on the anisotropic Rayleigh spheres is enhanced at the electric dipole resonance, and may be further increased by tuning the anisotropic parameters. On the contrary, the optical forces on the anisotropic spheres can be largely reduced for anisotropic spheres with electromagnetic transparency.
DOI
We investigate the optical forces on the radially anisotropic spheres from an incident plane wave based on our generalized full-wave scattering theory and the Maxwell stress tensor integration techniques. We demonstrate that the optical force on the Rayleigh sphere with radial anisotropy does not always obey the well-known Rayleigh’s law F~k40a6 (where k0 is the wave number and a is the radius of the sphere), but could be anomalous with the laws such as F~k00a2, F~k−20a0, and F~k80a10 under certain conditions. Therefore, the optical force on the anisotropic Rayleigh spheres is enhanced at the electric dipole resonance, and may be further increased by tuning the anisotropic parameters. On the contrary, the optical forces on the anisotropic spheres can be largely reduced for anisotropic spheres with electromagnetic transparency.
DOI
Monday, November 3, 2014
Squeezing red blood cells on an optical waveguide to monitor cell deformability during blood storage
Balpreet Singh Ahluwalia, Peter McCourt, Ana Oteiza, James S. Wilkinson, Thomas R Huser and Olav Gauta Helleso
Red blood cells squeeze through micro-capillaries as part of blood circulation in the body. The deformability of red blood cells is thus critical for blood circulation. In this work, we report a method to optically squeeze red blood cells using the evanescent field present on top of a planar waveguide chip. The optical forces from a narrow waveguide are used to squeeze red blood cells to a size comparable to the waveguide width. Optical forces and pressure distributions on the cells are numerically computed to explain the squeezing process. The proposed technique is used to quantify the loss of blood deformability that occurs during blood storage lesion. Squeezing red blood cells using waveguides is a sensitive technique and works simultaneously on several cells, making the method suitable for monitoring stored blood.
DOI
Red blood cells squeeze through micro-capillaries as part of blood circulation in the body. The deformability of red blood cells is thus critical for blood circulation. In this work, we report a method to optically squeeze red blood cells using the evanescent field present on top of a planar waveguide chip. The optical forces from a narrow waveguide are used to squeeze red blood cells to a size comparable to the waveguide width. Optical forces and pressure distributions on the cells are numerically computed to explain the squeezing process. The proposed technique is used to quantify the loss of blood deformability that occurs during blood storage lesion. Squeezing red blood cells using waveguides is a sensitive technique and works simultaneously on several cells, making the method suitable for monitoring stored blood.
DOI
Separation of blood cells with differing deformability using deterministic lateral displacement
David Holmes, Graeme Whyte, Joe Bailey, Nuria Vergara-Irigaray, Andrew Ekpenyong, Jochen Guck and Tom Duke
Determining cell mechanical properties is increasingly recognized as a marker-free way to characterize and separate biological cells. This emerging realization has led to the development of a plethora of appropriate measurement techniques. Here, we use a fairly novel approach, deterministic lateral displacement (DLD), to separate blood cells based on their mechanical phenotype with high throughput. Human red blood cells were treated chemically to alter their membrane deformability and the effect of this alteration on the hydrodynamic behaviour of the cells in a DLD device was investigated. Cells of defined stiffness (glutaraldehyde cross-linked erythrocytes) were used to test the performance of the DLD device across a range of cell stiffness and applied shear rates. Optical stretching was used as an independent method for quantifying the variation in stiffness of the cells. Lateral displacement of cells flowing within the device, and their subsequent exit position from the device were shown to correlate with cell stiffness. Data showing how the isolation of leucocytes from whole blood varies with applied shear rate are also presented. The ability to sort leucocyte sub-populations (T-lymphocytes and neutrophils), based on a combination of cell size and deformability, demonstrates the potential for using DLD devices to perform continuous fractionation and/or enrichment of leucocyte sub-populations from whole blood.
DOI
Determining cell mechanical properties is increasingly recognized as a marker-free way to characterize and separate biological cells. This emerging realization has led to the development of a plethora of appropriate measurement techniques. Here, we use a fairly novel approach, deterministic lateral displacement (DLD), to separate blood cells based on their mechanical phenotype with high throughput. Human red blood cells were treated chemically to alter their membrane deformability and the effect of this alteration on the hydrodynamic behaviour of the cells in a DLD device was investigated. Cells of defined stiffness (glutaraldehyde cross-linked erythrocytes) were used to test the performance of the DLD device across a range of cell stiffness and applied shear rates. Optical stretching was used as an independent method for quantifying the variation in stiffness of the cells. Lateral displacement of cells flowing within the device, and their subsequent exit position from the device were shown to correlate with cell stiffness. Data showing how the isolation of leucocytes from whole blood varies with applied shear rate are also presented. The ability to sort leucocyte sub-populations (T-lymphocytes and neutrophils), based on a combination of cell size and deformability, demonstrates the potential for using DLD devices to perform continuous fractionation and/or enrichment of leucocyte sub-populations from whole blood.
DOI
Microscale mapping of oscillatory flows
Spas Nedev, S. Carretero-Palacios, S. R. Kirchner, F. Jäckel and J. Feldmann
We present an optofluidic method that allows the two-dimensional vectorial near-field mapping of oscillatory flows with micron-scale resolution. An oscillatory flow created by a microsource (an optically trapped silica particle set to oscillate in a dipole-type mode) is detected by another twin silica particle independently trapped and located in the vicinity of the source. Fourier analysis of the motion of the detecting particle at different points in space and time renders the vectorial velocity map around the oscillating microsphere. The method introduced here paves the way for in-situ characterization of fast mixing microscale devices and for new detection methods able to provide location and recognition (due to the field pattern) of moving sources that may be applied to both artificial and living microobjects, including macromolecules, cells, and microorganisms.
DOI
We present an optofluidic method that allows the two-dimensional vectorial near-field mapping of oscillatory flows with micron-scale resolution. An oscillatory flow created by a microsource (an optically trapped silica particle set to oscillate in a dipole-type mode) is detected by another twin silica particle independently trapped and located in the vicinity of the source. Fourier analysis of the motion of the detecting particle at different points in space and time renders the vectorial velocity map around the oscillating microsphere. The method introduced here paves the way for in-situ characterization of fast mixing microscale devices and for new detection methods able to provide location and recognition (due to the field pattern) of moving sources that may be applied to both artificial and living microobjects, including macromolecules, cells, and microorganisms.
DOI
Tailoring azimuthal optical force on lossy chiral particles in Bessel beams
Huajin Chen, Neng Wang, Wanli Lu, Shiyang Liu, and Zhifang Lin
Based on the Mie scattering theory and Maxwell stress tensor method, we investigate the transverse optical force (TOF) acting on chiral particles illuminated by a zero-order Bessel beam. It is demonstrated that the particle chirality can induce an azimuthal optical force (AOF), resulting in orbital motion of particles around the optical beam axis. The AOF depends strongly on particle loss as well as the handedness of chirality, with its amplitude capable of changing by over an order of magnitude by particle's chiral loss. The other component of TOF, the radial optical force (ROF), is much less sensitive to the magnitude and handedness of the particle chirality as well as the loss when the chirality is small. Analytical result based on dipole approximation reveals that the AOF arises from the direct coupling of particle chirality to both the spin angular momentum (SAM) and optical vorticity (curl of Poynting vector), exhibiting a conversion of optical SAM of an incident beam to mechanical orbital angular momentum of an illuminated particle. Differently, the ROF originates from the transverse gradient force. In addition, particle chirality yields a negative contribution to the gradient force; thus the ROF can be attenuated and even reversed in direction when particle chirality is sufficiently large. These characteristics of TOF might find applications in chirality detection as well as sorting chiral particles of different handedness and separating them from conventional ones.
DOI
Based on the Mie scattering theory and Maxwell stress tensor method, we investigate the transverse optical force (TOF) acting on chiral particles illuminated by a zero-order Bessel beam. It is demonstrated that the particle chirality can induce an azimuthal optical force (AOF), resulting in orbital motion of particles around the optical beam axis. The AOF depends strongly on particle loss as well as the handedness of chirality, with its amplitude capable of changing by over an order of magnitude by particle's chiral loss. The other component of TOF, the radial optical force (ROF), is much less sensitive to the magnitude and handedness of the particle chirality as well as the loss when the chirality is small. Analytical result based on dipole approximation reveals that the AOF arises from the direct coupling of particle chirality to both the spin angular momentum (SAM) and optical vorticity (curl of Poynting vector), exhibiting a conversion of optical SAM of an incident beam to mechanical orbital angular momentum of an illuminated particle. Differently, the ROF originates from the transverse gradient force. In addition, particle chirality yields a negative contribution to the gradient force; thus the ROF can be attenuated and even reversed in direction when particle chirality is sufficiently large. These characteristics of TOF might find applications in chirality detection as well as sorting chiral particles of different handedness and separating them from conventional ones.
DOI
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