Oxidative stress is encountered in many biological systems; the resultant oxidative injury plays a significant role in the pathogenesis of diverse diseases. Conventional measurements on oxidative injury are employed almost exclusively on a large population of cells either by counting the fraction of cell death or by observing the fluorometric change resulting from exogenous reagents, thereby lacking in molecular detail and temporal specificity. In this work we combine laser tweezers and Raman spectroscopy to observe the response of single cells to oxidative stress. By measuring the temporal changes of vibrational spectra of single optically trapped cells, we demonstrate a molecular-level assessment of cellular oxidative injury in real time, both qualitatively and quantitatively, without the introduction of exogenous reagents. The main experimental findings are supported by the observation of Raman spectra of intermediates and downstream products. The abrogation of the above changes by ascorbic acid further illustrates the therapeutic effect of antioxidants against cellular oxidative injury. This approach is extensible to studies exploring the biochemical transformation of single cells or intracellular organelles in response to various chemical or physical stimuli. With the aid of molecular fingerprints, single-cell Raman spectroscopy exhibits a great potential for accessing the chemical aspects of cellular bioactivity, yielding insight into pathophysiological processes and assisting the development of novel therapeutic interventions against diseases.
Friday, October 23, 2009
Real-time molecular assessment on oxidative injury of single cells using Raman spectroscopy
Wei-Tien Chang, Hung-Lung Lin, Hung-Che Chen, Yao-Ming Wu, Wen-Jone Chen, Yuan-Teh Lee, Ian Liau
Oxidative stress is encountered in many biological systems; the resultant oxidative injury plays a significant role in the pathogenesis of diverse diseases. Conventional measurements on oxidative injury are employed almost exclusively on a large population of cells either by counting the fraction of cell death or by observing the fluorometric change resulting from exogenous reagents, thereby lacking in molecular detail and temporal specificity. In this work we combine laser tweezers and Raman spectroscopy to observe the response of single cells to oxidative stress. By measuring the temporal changes of vibrational spectra of single optically trapped cells, we demonstrate a molecular-level assessment of cellular oxidative injury in real time, both qualitatively and quantitatively, without the introduction of exogenous reagents. The main experimental findings are supported by the observation of Raman spectra of intermediates and downstream products. The abrogation of the above changes by ascorbic acid further illustrates the therapeutic effect of antioxidants against cellular oxidative injury. This approach is extensible to studies exploring the biochemical transformation of single cells or intracellular organelles in response to various chemical or physical stimuli. With the aid of molecular fingerprints, single-cell Raman spectroscopy exhibits a great potential for accessing the chemical aspects of cellular bioactivity, yielding insight into pathophysiological processes and assisting the development of novel therapeutic interventions against diseases.
Oxidative stress is encountered in many biological systems; the resultant oxidative injury plays a significant role in the pathogenesis of diverse diseases. Conventional measurements on oxidative injury are employed almost exclusively on a large population of cells either by counting the fraction of cell death or by observing the fluorometric change resulting from exogenous reagents, thereby lacking in molecular detail and temporal specificity. In this work we combine laser tweezers and Raman spectroscopy to observe the response of single cells to oxidative stress. By measuring the temporal changes of vibrational spectra of single optically trapped cells, we demonstrate a molecular-level assessment of cellular oxidative injury in real time, both qualitatively and quantitatively, without the introduction of exogenous reagents. The main experimental findings are supported by the observation of Raman spectra of intermediates and downstream products. The abrogation of the above changes by ascorbic acid further illustrates the therapeutic effect of antioxidants against cellular oxidative injury. This approach is extensible to studies exploring the biochemical transformation of single cells or intracellular organelles in response to various chemical or physical stimuli. With the aid of molecular fingerprints, single-cell Raman spectroscopy exhibits a great potential for accessing the chemical aspects of cellular bioactivity, yielding insight into pathophysiological processes and assisting the development of novel therapeutic interventions against diseases.
Emergence of exotic spatio-temporal structure under photon flux
K Yoshikawa
The operation of laser focusing on an object implies the creation of dielectric dielectric field under a thermodynamically open condition. Thus, we can expect the appearance of the effect of thermal irreversibility, such as breakdown of detailed balance, occurrence of circular state-flux in the phase-space, and limit-cycle oscillation. In the present article, we describe our recent experimental results on various kinds of exotic time-dependent phenomena induced by the continuous irradiation of laser. 1) Generation/annihilation of droplets from binary homogeneous liquid induced by laser: It will be shown that focused laser induces micro-phase separation on an oil/water isotropic solution. By choosing the proper experimental conditions, rhythmic change of generation, growth, and disappearance of a droplet at the focus is generated. This rhythmic phenomenon is a kind of limit-cycle oscillation. 2) Positive/negative photophoresis on a droplet: We show that a droplet is driven by a laser beam, either toward and backward along the direction of photon flux, through the change of the position of irradiation. Such photophoretic motion is induced by interfacial instability owe to the laser irradiation. 3) Rhythmic growth and bursting of a cluster with micro-beads: It is shown that negatively charged micro-beads are collected toward the focus of IR laser, i.e., optical tweezers. When the focusing angle is decreased from usual conditions, rhythmic change of the formation-growth-bursting of the beads cluster is generated.
Conservative and Nonconservative Torques in Optical Binding
D. Haefner, S. Sukhov, and A. Dogariu
We show that in the canonical case of two lossless spheres that are electromagnetically coupled there is interplay between conservative and nonconservative forces, which is controlled by the polarization of the bounding field. We demonstrate that this phenomenon leads to new mechanisms to induce torques on spherically symmetric, optically isotropic, and lossless objects. The electromagnetic interaction can be exploited to apply orbital torque about the mutual center of mass of thebounded spheres as well as spin around the individual axes. When the incident field is linearly polarized, the torques are mostly conservative and affect only transient behaviors while for circularly polarized fields, the torques are entirely nonconservative, resulting in steady rotations. Means to control the magnitudes of orbital and spin torques are presented and applications to nanorotator machines are discussed.
We show that in the canonical case of two lossless spheres that are electromagnetically coupled there is interplay between conservative and nonconservative forces, which is controlled by the polarization of the bounding field. We demonstrate that this phenomenon leads to new mechanisms to induce torques on spherically symmetric, optically isotropic, and lossless objects. The electromagnetic interaction can be exploited to apply orbital torque about the mutual center of mass of thebounded spheres as well as spin around the individual axes. When the incident field is linearly polarized, the torques are mostly conservative and affect only transient behaviors while for circularly polarized fields, the torques are entirely nonconservative, resulting in steady rotations. Means to control the magnitudes of orbital and spin torques are presented and applications to nanorotator machines are discussed.
Wednesday, October 21, 2009
Lithium niobate nanowires synthesis, optical properties, and manipulation
Rachel Grange, Jae-Woo Choi, Chia-Lung Hsieh, Ye Pu, Arnaud Magrez, Rita Smajda, László Forró, and Demetri Psaltis
Free-standing lithium niobate nanowires (LiNbO3) are synthesized by the hydrothermal route. The polarization response of the second harmonic generation (SHG) signal is measured in a single nanowire and used to identify the crystal orientation by matching with bulk LiNbO3 nonlinear optical susceptibility. The electrical manipulation of a LiNbO3 nanowire and its monitoring through the SHG signal in a fluidic setup are demonstrated.
Free-standing lithium niobate nanowires (LiNbO3) are synthesized by the hydrothermal route. The polarization response of the second harmonic generation (SHG) signal is measured in a single nanowire and used to identify the crystal orientation by matching with bulk LiNbO3 nonlinear optical susceptibility. The electrical manipulation of a LiNbO3 nanowire and its monitoring through the SHG signal in a fluidic setup are demonstrated.
Polarization Raman study of protein ordering by controllable RBC deformation
Satish Rao, Stefan Bálint, Luisa del Carmen Frias, Dmitri Petrov
Polarized Raman spectroscopy is used to provide evidence of hemoglobin protein ordering as the red blood cell (RBC) is stretched with optical tweezers. The stretching of the cell is intended to mimic the deformation that it experiences as it travels through vessels and capillaries. The depolarization ratios for a number of heme Raman bands change as the cell is stretched, confirming the semi-ordered nature of the hemoglobin ensemble in the cytoplasm. Furthermore, trends observed in the ratio shifts point to increased packing and ordering of the Hb after cell stretching. This evidence should shed more light on to the role of deformation in the RBC function.
Polarized Raman spectroscopy is used to provide evidence of hemoglobin protein ordering as the red blood cell (RBC) is stretched with optical tweezers. The stretching of the cell is intended to mimic the deformation that it experiences as it travels through vessels and capillaries. The depolarization ratios for a number of heme Raman bands change as the cell is stretched, confirming the semi-ordered nature of the hemoglobin ensemble in the cytoplasm. Furthermore, trends observed in the ratio shifts point to increased packing and ordering of the Hb after cell stretching. This evidence should shed more light on to the role of deformation in the RBC function.
Giant Enhanced Diffusion of Gold Nanoparticles in Optical Vortex Fields
Silvia Albaladejo, Manuel I. Marqués, Frank Scheffold and Juan Jose Saenz
We study the diffusion of a metal nanoparticle in the nonconservative force field of an optical vertex lattice. Radiation in the vortex array is shown to induce a giant enhancement over the free thermal diffusion. Langevin dynamics simulations show that the diffusion coefficient of (50 nm radius) gold particles at room temperature is enhanced by 2 orders of magnitude at power densities of the order or smaller than those used to trap nanoparticles with optical tweezers.
Spontaneous Oscillations of a Minimal Actomyosin System under Elastic Loading
P.-Y. Plaçais, M. Balland, T. Guérin, J.-F. Joanny, and P. Martin
Spontaneous mechanical oscillations occur in various types of biological systems where groups of motor molecules are elastically coupled to their environment. By using an optical trap to oppose the gliding motion of a single bead-tailed actin filament over a substrate densely coated with myosin motors, we mimicked this condition in vitro. We show that this minimal actomyosin system can oscillate spontaneously. Our finding accords quantitatively with a general theoretical framework where oscillatory instabilities emerge generically from the collective dynamics of molecular motors under load.
Spontaneous mechanical oscillations occur in various types of biological systems where groups of motor molecules are elastically coupled to their environment. By using an optical trap to oppose the gliding motion of a single bead-tailed actin filament over a substrate densely coated with myosin motors, we mimicked this condition in vitro. We show that this minimal actomyosin system can oscillate spontaneously. Our finding accords quantitatively with a general theoretical framework where oscillatory instabilities emerge generically from the collective dynamics of molecular motors under load.
Monday, October 19, 2009
History force on coated microbubbles propelled by ultrasound
Valeria Garbin, Benjamin Dollet, Marlies Overvelde, Dan Cojoc, Enzo Di Fabrizio, Leen van Wijngaarden, Andrea Prosperetti, Nico de Jong, Detlef Lohse, and Michel Versluis
In this paper the unsteady translation of coated microbubbles propelled by acoustic radiation force is studied experimentally. A system of two pulsating microbubbles of the type used as contrast agent in ultrasound medical imaging is considered, which attract each other as a result of the secondary Bjerknes force. Optical tweezers are used to isolate the bubble pair from neighboring boundaries so that it can be regarded as if in an unbounded fluid and the hydrodynamic forces acting on the system can be identified unambiguously. The radial and translational dynamics, excited by a 2.25 MHz ultrasound wave, is recorded with an ultrahigh speed camera at 15×106 frames/s. The time-resolved measurements reveal a quasisteady component of the translational velocity, at an average translational Reynolds number 0.5, and an oscillatory component at the same frequency as the radial pulsations, as predicted by existing models. Since the coating enforces a no-slip boundary condition, an increased viscous dissipation is expected due to the oscillatory component, similar to the case of an oscillating rigid sphere that was firstdescribed by Stokes [“On the effect of the internal friction of fluids on the motion of pendulums,” Trans. Cambridge Philos. Soc. 9, 8 (1851)]. A history force term is thereforeincluded in the force balance, in the form originally proposed by Basset and extended to the case of time-dependent radius by Takemura and Magnaudet [“The history force on a rapidly shrinking bubble rising at finite Reynolds number,” Phys. Fluids 16, 3247 (2004)]. The instantaneous values of the hydrodynamic forces extracted from the experimental data confirm that the history force accounts for the largest part of the viscous force. The trajectories of the bubbles predicted by numerically solving the equations of motion are in very good agreement with the experiment.
DOI
In this paper the unsteady translation of coated microbubbles propelled by acoustic radiation force is studied experimentally. A system of two pulsating microbubbles of the type used as contrast agent in ultrasound medical imaging is considered, which attract each other as a result of the secondary Bjerknes force. Optical tweezers are used to isolate the bubble pair from neighboring boundaries so that it can be regarded as if in an unbounded fluid and the hydrodynamic forces acting on the system can be identified unambiguously. The radial and translational dynamics, excited by a 2.25 MHz ultrasound wave, is recorded with an ultrahigh speed camera at 15×106 frames/s. The time-resolved measurements reveal a quasisteady component of the translational velocity, at an average translational Reynolds number 0.5, and an oscillatory component at the same frequency as the radial pulsations, as predicted by existing models. Since the coating enforces a no-slip boundary condition, an increased viscous dissipation is expected due to the oscillatory component, similar to the case of an oscillating rigid sphere that was firstdescribed by Stokes [“On the effect of the internal friction of fluids on the motion of pendulums,” Trans. Cambridge Philos. Soc. 9, 8 (1851)]. A history force term is thereforeincluded in the force balance, in the form originally proposed by Basset and extended to the case of time-dependent radius by Takemura and Magnaudet [“The history force on a rapidly shrinking bubble rising at finite Reynolds number,” Phys. Fluids 16, 3247 (2004)]. The instantaneous values of the hydrodynamic forces extracted from the experimental data confirm that the history force accounts for the largest part of the viscous force. The trajectories of the bubbles predicted by numerically solving the equations of motion are in very good agreement with the experiment.
DOI
Optical manipulation of proteins in aqueous solution
Yasuyuki Tsuboi, Tatsuya Shoji, Masayuki Nishino, Seiji Masuda, Koichiro Ishimori and Noboru Kitamura
Optical trapping of lysozyme, cytochrome c, or myoglobin based on photon pressure generated by focusing 1064 nm laser beam in an aqueous solution was explored. For all the proteins, microparticle formation was observed at the focal point under an optical microscope. Furthermore, the microparticles were identified to the molecular assemblies of the corresponding protein by means of confocal Raman microspectroscopy. For lysozyme, molecular clusters in solution were optically trapped to form the microparticle and it took more than 1 h to produce the microparticle. By contrast, molecular assembling proceeded within 1 min for cytochrome c and myoglobin. Since heme in cytochrome c or myoglobin would have a high polarizability, that would contribute to rapid assembling of the protein. Thus we demonstrated that a focused laser beam was a powerful tool to manipulate protein molecules in solution.
Optical trapping of lysozyme, cytochrome c, or myoglobin based on photon pressure generated by focusing 1064 nm laser beam in an aqueous solution was explored. For all the proteins, microparticle formation was observed at the focal point under an optical microscope. Furthermore, the microparticles were identified to the molecular assemblies of the corresponding protein by means of confocal Raman microspectroscopy. For lysozyme, molecular clusters in solution were optically trapped to form the microparticle and it took more than 1 h to produce the microparticle. By contrast, molecular assembling proceeded within 1 min for cytochrome c and myoglobin. Since heme in cytochrome c or myoglobin would have a high polarizability, that would contribute to rapid assembling of the protein. Thus we demonstrated that a focused laser beam was a powerful tool to manipulate protein molecules in solution.
Friday, October 16, 2009
Simple Route for Preparing Optically Trappable Probes for Surface-Enhanced Raman Scattering
Stefan Balint, Mark P. Kreuzer, Satish Rao, Gonal Badenes, Pavol Mikovsk and Dmitri Petrov
Surface-enhanced Raman scattering (SERS) has proven to be a powerful technique for a wide range of topics such as chemical analysis, materials research, and biological imaging. In this work, we report in detail on the development and characterization of micrometer-sized dielectric beads with metal colloids attached to their surface. The metalized beads were sufficiently transparent enabling optical trapping while the presence of metal islands provided the SERS. This method is fast and simple and is void of complications when compared to the Tollen’s test used in previous publications. This highly efficient probe can be placed and scanned with nanometric accuracy near living cells. The process to create such probes is described including discussion of various parameters that are critical in achieving the desired results. Additionally, their use is demonstrated by detecting low quantities of a drug in aqueous solution and in a cell membrane.
Surface-enhanced Raman scattering (SERS) has proven to be a powerful technique for a wide range of topics such as chemical analysis, materials research, and biological imaging. In this work, we report in detail on the development and characterization of micrometer-sized dielectric beads with metal colloids attached to their surface. The metalized beads were sufficiently transparent enabling optical trapping while the presence of metal islands provided the SERS. This method is fast and simple and is void of complications when compared to the Tollen’s test used in previous publications. This highly efficient probe can be placed and scanned with nanometric accuracy near living cells. The process to create such probes is described including discussion of various parameters that are critical in achieving the desired results. Additionally, their use is demonstrated by detecting low quantities of a drug in aqueous solution and in a cell membrane.
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