Lei Shao, Zhong-Jian Yang, Daniel Andrén, Peter Johansson, and Mikael Käll
Efficient and robust artificial nanomotors could provide a variety of exciting possibilities for applications in physics, biology and chemistry, including nanoelectromechanical systems, biochemical sensing, and drug delivery. However, the application of current man-made nanomotors is limited by their sophisticated fabrication techniques, low mechanical output power and severe environmental requirements, making their performance far below that of natural biomotors. Here we show that single-crystal gold nanorods can be rotated extremely fast in aqueous solutions through optical torques dominated by plasmonic resonant scattering of circularly polarized laser light with power as low as a few mW. The nanorods are trapped in 2D against a glass surface, and their rotational dynamics is highly dependent on their surface plasmon resonance properties. They can be kept continuously rotating for hours with limited photothermal side effects and they can be applied for detection of molecular binding with high sensitivity. Because of their biocompatibility, mechanical and thermal stability, and record rotation speeds reaching up to 42 kHz (2.5 million revolutions per minute), these rotary nanomotors could advance technologies to meet a wide range of future nanomechanical and biomedical needs in fields such as nanorobotics, nanosurgery, DNA manipulation and nano/microfluidic flow control.
DOI
Wednesday, November 18, 2015
Dynamic chromatin regulation from a single molecule perspective
Beat Fierz
Chromatin regulatory processes, like all biological reactions, are dynamic and stochastic in nature, but can give rise to stable and inheritable changes in gene expression patterns. A molecular understanding of those processes is key for fundamental biological insight into gene regulation, epigenetic inheritance, lineage determination and therapeutic intervention in the case of disease. In recent years great progress has been made in identifying important molecular players involved in key chromatin regulatory pathways. Conversely, we are only beginning to understand the dynamic interplay between protein effectors, transcription factors and the chromatin substrate itself. Single-molecule approaches employing both highly defined chromatin substrates in vitro, as well as direct observation of complex regulatory processes in vivo open new avenues for a molecular view of chromatin regulation. This review highlights recent applications of single-molecule methods and related techniques to investigate fundamental chromatin regulatory processes.
DOI
Chromatin regulatory processes, like all biological reactions, are dynamic and stochastic in nature, but can give rise to stable and inheritable changes in gene expression patterns. A molecular understanding of those processes is key for fundamental biological insight into gene regulation, epigenetic inheritance, lineage determination and therapeutic intervention in the case of disease. In recent years great progress has been made in identifying important molecular players involved in key chromatin regulatory pathways. Conversely, we are only beginning to understand the dynamic interplay between protein effectors, transcription factors and the chromatin substrate itself. Single-molecule approaches employing both highly defined chromatin substrates in vitro, as well as direct observation of complex regulatory processes in vivo open new avenues for a molecular view of chromatin regulation. This review highlights recent applications of single-molecule methods and related techniques to investigate fundamental chromatin regulatory processes.
DOI
Lateral optical force on paired chiral nanoparticles in linearly polarized plane waves
Huajin Chen, Yikun Jiang, Neng Wang, Wanli Lu, Shiyang Liu, and Zhifang Lin
We demonstrate that a lateral optical force (LOF) can be induced on paired chiral nanoparticles with opposite handedness under the illumination of a linearly polarized plane wave. The LOFs on both chiral particles are equal and thus can move the pair sideways, with the direction depending on the separation between two particles, as well as the handedness of particle chirality. Analytical theory reveals that the LOF comes largely from the optical potential gradient established by the multiple scattering of light between the paired particles with asymmetric chirality. In addition, it is weakly dependent on the material loss of a particle, a feature of gradient force, while heavily dependent on the magnitude and handedness of particle chirality. The effect is expected to find applications in sorting and separating chiral dimers of different handedness.
DOI
We demonstrate that a lateral optical force (LOF) can be induced on paired chiral nanoparticles with opposite handedness under the illumination of a linearly polarized plane wave. The LOFs on both chiral particles are equal and thus can move the pair sideways, with the direction depending on the separation between two particles, as well as the handedness of particle chirality. Analytical theory reveals that the LOF comes largely from the optical potential gradient established by the multiple scattering of light between the paired particles with asymmetric chirality. In addition, it is weakly dependent on the material loss of a particle, a feature of gradient force, while heavily dependent on the magnitude and handedness of particle chirality. The effect is expected to find applications in sorting and separating chiral dimers of different handedness.
DOI
Tuesday, November 17, 2015
In situ calibrating optical tweezers with sinusoidal-wave drag force method
Li Di, Zhou Jin-Hua, Hu Xin-Yao, Zhong Min-Cheng, Gong Lei, Wang Zi-Qiang, Wang Hao-Wei and Li Yin-Mei
We introduce a corrected sinusoidal-wave drag force method (SDFM) into optical tweezers to calibrate the trapping stiffness of the optical trap and conversion factor (CF) of photodetectors. First, the theoretical analysis and experimental result demonstrate that the correction of SDFM is necessary, especially the error of no correction is up to 11.25% for a bead of 5 μm in diameter. Second, the simulation results demonstrate that the SDFM has a better performance in the calibration of optical tweezers than the triangular-wave drag force method (TDFM) and power spectrum density method (PSDM) at the same signal-to-noise ratio or trapping stiffness. Third, in experiments, the experimental standard deviations of calibration of trapping stiffness and CF with the SDFM are about less than 50% of TDFM and PSDM especially at low laser power. Finally, the experiments of stretching DNA verify that the in situ calibration with the SDFM improves the measurement stability and accuracy.
DOI
We introduce a corrected sinusoidal-wave drag force method (SDFM) into optical tweezers to calibrate the trapping stiffness of the optical trap and conversion factor (CF) of photodetectors. First, the theoretical analysis and experimental result demonstrate that the correction of SDFM is necessary, especially the error of no correction is up to 11.25% for a bead of 5 μm in diameter. Second, the simulation results demonstrate that the SDFM has a better performance in the calibration of optical tweezers than the triangular-wave drag force method (TDFM) and power spectrum density method (PSDM) at the same signal-to-noise ratio or trapping stiffness. Third, in experiments, the experimental standard deviations of calibration of trapping stiffness and CF with the SDFM are about less than 50% of TDFM and PSDM especially at low laser power. Finally, the experiments of stretching DNA verify that the in situ calibration with the SDFM improves the measurement stability and accuracy.
DOI
Spectrally reconfigurable integrated multi-spot particle trap
Kaelyn D. Leake, Michael A. B. Olson, Damla Ozcelik, Aaron R. Hawkins, and Holger Schmidt
Optical manipulation of small particles in the form of trapping, pushing, or sorting has developed into a vast field with applications in the life sciences, biophysics, and atomic physics. Recently, there has been increasing effort toward integration of particle manipulation techniques with integrated photonic structures on self-contained optofluidic chips. Here, we use the wavelength dependence of multi-spot pattern formation in multimode interference (MMI) waveguides to create a new type of reconfigurable, integrated optical particle trap. Interfering lateral MMI modes create multiple trapping spots in an intersecting fluidic channel. The number of trapping spots can be dynamically controlled by altering the trapping wavelength. This novel, spectral reconfigurability is utilized to deterministically move single and multiple particles between different trapping locations along the channel. This fully integrated multi-particle trap can form the basis of high throughput biophotonic assays on a chip.
DOI
Optical manipulation of small particles in the form of trapping, pushing, or sorting has developed into a vast field with applications in the life sciences, biophysics, and atomic physics. Recently, there has been increasing effort toward integration of particle manipulation techniques with integrated photonic structures on self-contained optofluidic chips. Here, we use the wavelength dependence of multi-spot pattern formation in multimode interference (MMI) waveguides to create a new type of reconfigurable, integrated optical particle trap. Interfering lateral MMI modes create multiple trapping spots in an intersecting fluidic channel. The number of trapping spots can be dynamically controlled by altering the trapping wavelength. This novel, spectral reconfigurability is utilized to deterministically move single and multiple particles between different trapping locations along the channel. This fully integrated multi-particle trap can form the basis of high throughput biophotonic assays on a chip.
DOI
High-throughput linear optical stretcher for mechanical characterization of blood cells
Kevin B. Roth, Keith B. Neeves, Jeff Squier and David W. M. Marr
This study describes a linear optical stretcher as a high-throughput mechanical property cytometer. Custom, inexpensive, and scalable optics image a linear diode bar source into a microfluidic channel, where cells are hydrodynamically focused into the optical stretcher. Upon entering the stretching region, antipodal optical forces generated by the refraction of tightly focused laser light at the cell membrane deform each cell in flow. Each cell relaxes as it flows out of the trap and is compared to the stretched state to determine deformation. The deformation response of untreated red blood cells and neutrophils were compared to chemically treated cells. Statistically significant differences were observed between normal, diamide-treated, and glutaraldehyde-treated red blood cells, as well as between normal and cytochalasin D-treated neutrophils. Based on the behavior of the pure, untreated populations of red cells and neutrophils, a mixed population of these cells was tested and the discrete populations were identified by deformability.
DOI
This study describes a linear optical stretcher as a high-throughput mechanical property cytometer. Custom, inexpensive, and scalable optics image a linear diode bar source into a microfluidic channel, where cells are hydrodynamically focused into the optical stretcher. Upon entering the stretching region, antipodal optical forces generated by the refraction of tightly focused laser light at the cell membrane deform each cell in flow. Each cell relaxes as it flows out of the trap and is compared to the stretched state to determine deformation. The deformation response of untreated red blood cells and neutrophils were compared to chemically treated cells. Statistically significant differences were observed between normal, diamide-treated, and glutaraldehyde-treated red blood cells, as well as between normal and cytochalasin D-treated neutrophils. Based on the behavior of the pure, untreated populations of red cells and neutrophils, a mixed population of these cells was tested and the discrete populations were identified by deformability.
DOI
Monday, November 16, 2015
Diffusion and Reactivity in Ultraviscous Aerosol and the Correlation with Particle Viscosity
Frances H. Marshall, Rachael E.H. Miles, Young-Chul Song, Peter B. Ohm, Rory M. Power, Jonathan P Reid and Cari S. Dutcher
The slow transport of water, organic species and oxidants in viscous aerosol can lead to aerosol existing in transient states that are not solely governed by thermodynamic principles but by the kinetics of gas-particle partitioning. The relationship between molecular diffusion constants and particle viscosity (for example, as reflected in the Stokes-Einstein equation) is frequently considered to provide an approximate guide to relate the kinetics of aerosol transformation with a material property of the aerosol. We report direct studies of both molecular diffusion and viscosity in the aerosol phase for the ternary system water/maleic acid/sucrose, considering the relationship between the hygroscopic response associated with the change in water partitioning, the volatilisation of maleic acid, the ozonolysis kinetics of maleic acid and the particle viscosity. Although water clearly acts as a plasticiser, the addition of minor fractions of other organic moieties can similarly lead to significant changes in the viscosity from that expected for the dominant component forming the organic matrix (sucrose). Here we highlight that the Stokes-Einstein relationship between the diffusion constant of water and the viscosity of the particle may be more than an order of magnitude in error, even at viscosities as low as 1 Pa s. We show that the thermodynamic relationships of hygroscopic response that underpin such kinetic determinations must be accurately known to retrieve accurate values for diffusion constants; such data are often not available. Further, we show that scaling of the diffusion constants of organic molecules of similar size to those forming the matrix with particle viscosity may be well represented by the Stokes-Einstein equation, suppressing the apparent volatility of semi-volatile components. Finally, the variation in uptake coefficients and diffusion constants for oxidants and small weakly interacting molecules may be much less dependent on viscosity than the diffusion constants of more strongly interacting molecules such as water.
DOI
The slow transport of water, organic species and oxidants in viscous aerosol can lead to aerosol existing in transient states that are not solely governed by thermodynamic principles but by the kinetics of gas-particle partitioning. The relationship between molecular diffusion constants and particle viscosity (for example, as reflected in the Stokes-Einstein equation) is frequently considered to provide an approximate guide to relate the kinetics of aerosol transformation with a material property of the aerosol. We report direct studies of both molecular diffusion and viscosity in the aerosol phase for the ternary system water/maleic acid/sucrose, considering the relationship between the hygroscopic response associated with the change in water partitioning, the volatilisation of maleic acid, the ozonolysis kinetics of maleic acid and the particle viscosity. Although water clearly acts as a plasticiser, the addition of minor fractions of other organic moieties can similarly lead to significant changes in the viscosity from that expected for the dominant component forming the organic matrix (sucrose). Here we highlight that the Stokes-Einstein relationship between the diffusion constant of water and the viscosity of the particle may be more than an order of magnitude in error, even at viscosities as low as 1 Pa s. We show that the thermodynamic relationships of hygroscopic response that underpin such kinetic determinations must be accurately known to retrieve accurate values for diffusion constants; such data are often not available. Further, we show that scaling of the diffusion constants of organic molecules of similar size to those forming the matrix with particle viscosity may be well represented by the Stokes-Einstein equation, suppressing the apparent volatility of semi-volatile components. Finally, the variation in uptake coefficients and diffusion constants for oxidants and small weakly interacting molecules may be much less dependent on viscosity than the diffusion constants of more strongly interacting molecules such as water.
DOI
Spatial Filtering of a Diode Laser Beam for Confocal Raman Microscopy
Kitt, Jay P.; Bryce, David A.; Harris, Joel M.
With the development of single-longitudinal mode diode lasers, there has been an increase in using these sources for Raman spectroscopy. This is largely due to the cost-effectiveness of diode lasers, which offer savings not only in initial capital cost, but also electrical, cooling, and replacement costs over time, when compared with ion lasers. The use of diode-lasers in confocal Raman microscopy has remained a challenge, however, due to poor transverse beam quality. In this work, we present the design and implementation of a simple spatial filter capable of adapting a single-mode diode laser source to confocal Raman microscopy, yielding comparable spatial resolution as a gas-ion laser beam for profiling and optical-trapping applications. For profiling applications, spatial filtering improved x,y resolution of the beam by a factor 10, which in turn increased optical-trapping forces by ∼90 times and yielded sevenfold greater Raman scattering signal intensity from an optically trapped phospholipid vesicle.
DOI
With the development of single-longitudinal mode diode lasers, there has been an increase in using these sources for Raman spectroscopy. This is largely due to the cost-effectiveness of diode lasers, which offer savings not only in initial capital cost, but also electrical, cooling, and replacement costs over time, when compared with ion lasers. The use of diode-lasers in confocal Raman microscopy has remained a challenge, however, due to poor transverse beam quality. In this work, we present the design and implementation of a simple spatial filter capable of adapting a single-mode diode laser source to confocal Raman microscopy, yielding comparable spatial resolution as a gas-ion laser beam for profiling and optical-trapping applications. For profiling applications, spatial filtering improved x,y resolution of the beam by a factor 10, which in turn increased optical-trapping forces by ∼90 times and yielded sevenfold greater Raman scattering signal intensity from an optically trapped phospholipid vesicle.
DOI
Friday, November 13, 2015
Nanoscale volume confinement and fluorescence enhancement with double nanohole aperture
Raju Regmi, Ahmed A. Al Balushi, Hervé Rigneault, Reuven Gordon & Jérôme Wenger
Diffraction ultimately limits the fluorescence collected from a single molecule, and sets an upper limit to the maximum concentration to isolate a single molecule in the detection volume. To overcome these limitations, we introduce here the use of a double nanohole structure with 25 nm gap, and report enhanced detection of single fluorescent molecules in concentrated solutions exceeding 20 micromolar. The nanometer gap concentrates the light into an apex volume down to 70 zeptoliter (10−21 L), 7000-fold below the diffraction-limited confocal volume. Using fluorescence correlation spectroscopy and time-correlated photon counting, we measure fluorescence enhancement up to 100-fold, together with local density of optical states (LDOS) enhancement of 30-fold. The distinctive features of double nanoholes combining high local field enhancement, efficient background screening and relative nanofabrication simplicity offer new strategies for real time investigation of biochemical events with single molecule resolution at high concentrations.
DOI
Diffraction ultimately limits the fluorescence collected from a single molecule, and sets an upper limit to the maximum concentration to isolate a single molecule in the detection volume. To overcome these limitations, we introduce here the use of a double nanohole structure with 25 nm gap, and report enhanced detection of single fluorescent molecules in concentrated solutions exceeding 20 micromolar. The nanometer gap concentrates the light into an apex volume down to 70 zeptoliter (10−21 L), 7000-fold below the diffraction-limited confocal volume. Using fluorescence correlation spectroscopy and time-correlated photon counting, we measure fluorescence enhancement up to 100-fold, together with local density of optical states (LDOS) enhancement of 30-fold. The distinctive features of double nanoholes combining high local field enhancement, efficient background screening and relative nanofabrication simplicity offer new strategies for real time investigation of biochemical events with single molecule resolution at high concentrations.
DOI
Enhanced Optical Manipulation of Cells Using Antireflection Coated Microparticles
Derek Craig, Alison McDonald, Michael Mazilu, Helen Rendall, Frank Gunn-Moore, and Kishan Dholakia
We demonstrate the use of antireflection (AR) coated microparticles for the enhanced optical manipulation of cells. Specifically, we incubate CHO-K1, HL60, and NMuMG cell lines with AR-coated titania microparticles and subsequently performed drag force measurements using optical trapping. Direct comparisons were performed between native, polystyrene microparticle and AR microparticle tagged cells. The optical trapping efficiency was recorded by measuring the Q value in a drag force experiment. CHO-K1 cells incubated with AR microparticles show an increase in the Q value of nearly 220% versus native cells. With the inclusion of AR microparticles, cell velocities exceeding 50 μm/s were recorded for only 33 mW of laser trapping power. Cell viability was confirmed with fluorescent dyes and cells expressing a fluorescent ubiquitination-based cell cycle protein (FUCCI), which verified no disruption to the cell cycle in the presence of AR microparticles.
DOI
We demonstrate the use of antireflection (AR) coated microparticles for the enhanced optical manipulation of cells. Specifically, we incubate CHO-K1, HL60, and NMuMG cell lines with AR-coated titania microparticles and subsequently performed drag force measurements using optical trapping. Direct comparisons were performed between native, polystyrene microparticle and AR microparticle tagged cells. The optical trapping efficiency was recorded by measuring the Q value in a drag force experiment. CHO-K1 cells incubated with AR microparticles show an increase in the Q value of nearly 220% versus native cells. With the inclusion of AR microparticles, cell velocities exceeding 50 μm/s were recorded for only 33 mW of laser trapping power. Cell viability was confirmed with fluorescent dyes and cells expressing a fluorescent ubiquitination-based cell cycle protein (FUCCI), which verified no disruption to the cell cycle in the presence of AR microparticles.
DOI
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