Paden B. Roder, Sandeep Manandhar, Bennett E. Smith, Xuezhe Zhou, Vaithiyalingam S. Shutthanandan and Peter J. Pauzauskie
Elucidating the temperatures of particles trapped in laser tweezers is of great importance and impacts studies and applications in various areas of science and engineering. A unique method is demonstrated for extracting the local temperature of trapped ion-implanted silicon nanowires undergoing hot Brownian motion in superheated water, and their efficacy as agents for photothermal cancer therapy is investigated.
DOI
Friday, November 13, 2015
Thursday, November 12, 2015
Automated analysis of single cells using Laser Tweezers Raman Spectroscopy
Stephen Casabella, Peter Gardner, Patricia Scully and N J Goddard
In recent years, significant progress has been made into the label-free detection and discrimination of individual cancer cells using Laser Tweezers Raman Spectroscopy (LTRS). However, the majority of examples reported have involved manual trapping of cells, which is time consuming and may lead to different cell lines being analysed in discrete batches. A simple, low-cost microfluidic flow chamber is introduced which allows single cells to be optically trapped and analysed in an automated fashion, greatly reducing the level of operator input required. Two implementations of the flow chamber are discussed here; a basic single-channel device in which the fluid velocity is controlled manually, and a dual-channel device which permits the automated capture and analysis of multiple cell lines with no operator input. Results are presented for the discrimination of live epithelial prostate cells and lymphocytes, together with a consideration of the consequences of traditional ‘batch analysis’ typically used for LTRS of live cells.
DOI
In recent years, significant progress has been made into the label-free detection and discrimination of individual cancer cells using Laser Tweezers Raman Spectroscopy (LTRS). However, the majority of examples reported have involved manual trapping of cells, which is time consuming and may lead to different cell lines being analysed in discrete batches. A simple, low-cost microfluidic flow chamber is introduced which allows single cells to be optically trapped and analysed in an automated fashion, greatly reducing the level of operator input required. Two implementations of the flow chamber are discussed here; a basic single-channel device in which the fluid velocity is controlled manually, and a dual-channel device which permits the automated capture and analysis of multiple cell lines with no operator input. Results are presented for the discrimination of live epithelial prostate cells and lymphocytes, together with a consideration of the consequences of traditional ‘batch analysis’ typically used for LTRS of live cells.
DOI
Recent advances in holographic 3D particle tracking
Pasquale Memmolo, Lisa Miccio, Melania Paturzo, Giuseppe Di Caprio, Giuseppe Coppola, Paolo A. Netti, and Pietro Ferraro
Particle tracking is a fundamental technique for investigating a variety of biophysical processes, from intracellular dynamics to the characterization of cell motility and migration. However, observing three-dimensional (3D) trajectories of particles is in general a challenging task in classical microscopy owing to the limited imaging depth of field of commercial optical microscopes, which represents a serious drawback for the analysis of time-lapse microscopy image data. Therefore, numerous automated particle-tracking approaches have been developed by many research groups around the world. Recently, digital holography (DH) in microscopy has rapidly gained credit as one of the elective techniques for these applications, mainly due to the uniqueness of the DH to provide a posteriori quantitative multiple refocusing capability and phase-contrast imaging. Starting from this paradigm, a huge amount of 3D holographic tracking approaches have been conceived and investigated for applications in various branches of science, including optofluids, microfluidics, biomedical microscopy, cell mechano-trasduction, and cell migration. Since a wider community of readers could be interested in such a review, i.e., not only scientists working in the fields of optics and photonics but also users of particle-tracking tools, it should be very beneficial to provide a complete review of state-of-the-art holographic 3D particle-tracking methods and their applications in bio-microfluidics.
DOI
Particle tracking is a fundamental technique for investigating a variety of biophysical processes, from intracellular dynamics to the characterization of cell motility and migration. However, observing three-dimensional (3D) trajectories of particles is in general a challenging task in classical microscopy owing to the limited imaging depth of field of commercial optical microscopes, which represents a serious drawback for the analysis of time-lapse microscopy image data. Therefore, numerous automated particle-tracking approaches have been developed by many research groups around the world. Recently, digital holography (DH) in microscopy has rapidly gained credit as one of the elective techniques for these applications, mainly due to the uniqueness of the DH to provide a posteriori quantitative multiple refocusing capability and phase-contrast imaging. Starting from this paradigm, a huge amount of 3D holographic tracking approaches have been conceived and investigated for applications in various branches of science, including optofluids, microfluidics, biomedical microscopy, cell mechano-trasduction, and cell migration. Since a wider community of readers could be interested in such a review, i.e., not only scientists working in the fields of optics and photonics but also users of particle-tracking tools, it should be very beneficial to provide a complete review of state-of-the-art holographic 3D particle-tracking methods and their applications in bio-microfluidics.
DOI
Wednesday, November 11, 2015
Developing a video tracking method to study interactions between close pairs of optically trapped particles in three dimensions
Allan Raudsepp, Marjorie Griffiths, Andrew J. Sutherland-Smith, and Martin A. K. Williams
We develop a video tracking method that utilizes an interpolation-based normalized cross-correlation approach to track the position of microscopic spherical particles in three dimensions. Subnanometer resolution is demonstrated. The method does not assume that the particle’s image is radially symmetric, making it useful for determining the position when particles are close and their images overlap. This is demonstrated in a study of the electrostatic and hydrodynamic interactions between a pair of beads in dual laser tweezers traps.
DOI
We develop a video tracking method that utilizes an interpolation-based normalized cross-correlation approach to track the position of microscopic spherical particles in three dimensions. Subnanometer resolution is demonstrated. The method does not assume that the particle’s image is radially symmetric, making it useful for determining the position when particles are close and their images overlap. This is demonstrated in a study of the electrostatic and hydrodynamic interactions between a pair of beads in dual laser tweezers traps.
DOI
On artifacts in single-molecule force spectroscopy
Pilar Cossioa, Gerhard Hummera, and Attila Szabo
In typical force spectroscopy experiments, a small biomolecule is attached to a soft polymer linker that is pulled with a relatively large bead or cantilever. At constant force, the total extension stochastically changes between two (or more) values, indicating that the biomolecule undergoes transitions between two (or several) conformational states. In this paper, we consider the influence of the dynamics of the linker and mesoscopic pulling device on the force-dependent rate of the conformational transition extracted from the time dependence of the total extension, and the distribution of rupture forces in force-clamp and force-ramp experiments, respectively. For these different experiments, we derive analytic expressions for the observables that account for the mechanical response and dynamics of the pulling device and linker. Possible artifacts arise when the characteristic times of the pulling device and linker become comparable to, or slower than, the lifetimes of the metastable conformational states, and when the highly anharmonic regime of stretched linkers is probed at high forces. We also revisit the problem of relating force-clamp and force-ramp experiments, and identify a linker and loading rate-dependent correction to the rates extracted from the latter. The theory provides a framework for both the design and the quantitative analysis of force spectroscopy experiments by highlighting, and correcting for, factors that complicate their interpretation.
DOI
In typical force spectroscopy experiments, a small biomolecule is attached to a soft polymer linker that is pulled with a relatively large bead or cantilever. At constant force, the total extension stochastically changes between two (or more) values, indicating that the biomolecule undergoes transitions between two (or several) conformational states. In this paper, we consider the influence of the dynamics of the linker and mesoscopic pulling device on the force-dependent rate of the conformational transition extracted from the time dependence of the total extension, and the distribution of rupture forces in force-clamp and force-ramp experiments, respectively. For these different experiments, we derive analytic expressions for the observables that account for the mechanical response and dynamics of the pulling device and linker. Possible artifacts arise when the characteristic times of the pulling device and linker become comparable to, or slower than, the lifetimes of the metastable conformational states, and when the highly anharmonic regime of stretched linkers is probed at high forces. We also revisit the problem of relating force-clamp and force-ramp experiments, and identify a linker and loading rate-dependent correction to the rates extracted from the latter. The theory provides a framework for both the design and the quantitative analysis of force spectroscopy experiments by highlighting, and correcting for, factors that complicate their interpretation.
DOI
Trapping and Manipulation of Copper Nanoparticles in Rayleigh Regime
E. Thanikaivelan, R. Jothilakshmi, P. Murugakoothan
The copper nanoparticles were synthesized by green technology using leaf extracts of Ocimum sanctum. The synthesized copper nanoparticles were confirmed by the change of colour after the addition of leaf extract into the copper sulfate solution. The synthesized copper was characterized by X-ray Diffraction (XRD), transmission electron microscopy (TEM) and UV-vis absorption spectroscopy. The copper nanoparticles are crystallized with FCC structure. The synthesized copper nanoparticles exhibit spherical morphology with average particle size of 20 nm. The copper nanoparticle exhibits absorption broad band between 550 nm – 575 nm. The optical trapping effect of Gaussian beam acting on a copper nanoparticle in Rayleigh regime was studied. The optical scattering and optical gradient forces were calculated for 20 nm copper particle.
DOI
The copper nanoparticles were synthesized by green technology using leaf extracts of Ocimum sanctum. The synthesized copper nanoparticles were confirmed by the change of colour after the addition of leaf extract into the copper sulfate solution. The synthesized copper was characterized by X-ray Diffraction (XRD), transmission electron microscopy (TEM) and UV-vis absorption spectroscopy. The copper nanoparticles are crystallized with FCC structure. The synthesized copper nanoparticles exhibit spherical morphology with average particle size of 20 nm. The copper nanoparticle exhibits absorption broad band between 550 nm – 575 nm. The optical trapping effect of Gaussian beam acting on a copper nanoparticle in Rayleigh regime was studied. The optical scattering and optical gradient forces were calculated for 20 nm copper particle.
DOI
Silver Nanoparticles - Trapping and Manipulation
E. Thanikaivalan, R. Jothilakshmi, P. Murugakoothan
Silver nanoparticles with different radii were synthesized using silver nitrate solution added with sodium borohydride solution at different ratios. The synthesized silver nanoparticles of radii 25.3 nm, 31 nm, 33.6 nm and 37.1 nm were characterized by transmission electron microscopy (TEM) and UV-vis absorption spectroscopy. The synthesized silver nanoparticles exhibit spherical morphology for all radii. The silver nanoparticles exhibit the plasmon resonance band between 390 nm – 400 nm. The optical trapping effect of Gaussian beam acting on a silver nanoparticle in Rayleigh regime was studied. The optical scattering and optical gradient forces were calculated for silver nanoparticles of different radii.
DOI
Silver nanoparticles with different radii were synthesized using silver nitrate solution added with sodium borohydride solution at different ratios. The synthesized silver nanoparticles of radii 25.3 nm, 31 nm, 33.6 nm and 37.1 nm were characterized by transmission electron microscopy (TEM) and UV-vis absorption spectroscopy. The synthesized silver nanoparticles exhibit spherical morphology for all radii. The silver nanoparticles exhibit the plasmon resonance band between 390 nm – 400 nm. The optical trapping effect of Gaussian beam acting on a silver nanoparticle in Rayleigh regime was studied. The optical scattering and optical gradient forces were calculated for silver nanoparticles of different radii.
DOI
Transverse Chiral Optical Forces by Chiral Surface Plasmon Polaritons
M.H. Alizadeh and Bjoern M. Reinhard
Recently the concepts of transverse spin angular momentum and Belinfante spin momentum of evanescent waves have drawn considerable attention. Here, we investigate these novel physical properties of electromagnetic fields in the context of chiral surface plasmon polaritons. We demonstrate, both analytically and numerically, that chiral surface plasmon polaritons possess transverse spin angular momentum and Belinfante momentum with rich and non-trivial characteristics. We also show that the transverse spin angular momentum of chiral surface plasmon polaritons leads to the emergence of transverse optical forces in opposite directions for chiral objects of different handedness. The magnitude of this chiral transverse optical force on a chiral particle is comparable to the magnitude of the achiral optical forces on the particle, namely the gradient force which arises from the intensity gradient and the scattering force which is the result of the linear momentum transfer of the photon to the particle. This finding may pave the way for realization of optical separation of chiral biomolecules.
DOI
Recently the concepts of transverse spin angular momentum and Belinfante spin momentum of evanescent waves have drawn considerable attention. Here, we investigate these novel physical properties of electromagnetic fields in the context of chiral surface plasmon polaritons. We demonstrate, both analytically and numerically, that chiral surface plasmon polaritons possess transverse spin angular momentum and Belinfante momentum with rich and non-trivial characteristics. We also show that the transverse spin angular momentum of chiral surface plasmon polaritons leads to the emergence of transverse optical forces in opposite directions for chiral objects of different handedness. The magnitude of this chiral transverse optical force on a chiral particle is comparable to the magnitude of the achiral optical forces on the particle, namely the gradient force which arises from the intensity gradient and the scattering force which is the result of the linear momentum transfer of the photon to the particle. This finding may pave the way for realization of optical separation of chiral biomolecules.
DOI
Tuesday, November 10, 2015
Single-molecule perspectives on helicase mechanisms and functions
Bo Sun & Michelle D. Wang
Helicases are a diverse group of molecular motors that utilize energy derived from the hydrolysis of nucleoside triphosphates (NTPs) to unwind and translocate along nucleic acids. These enzymes play critical roles in nearly all aspects of nucleic acid metabolism, and consequently, a detailed understanding of helicase mechanisms at the molecular level is essential. Over the past few decades, single-molecule techniques, such as optical tweezers, magnetic tweezers, laminar flow, fluorescence resonance energy transfer (FRET), and DNA curtains, have proved to be powerful tools to investigate the functional properties of both DNA and RNA helicases. These approaches allow researchers to manipulate single helicase molecules, perturb their free energy landscape to probe the chemo-mechanical activities of these motors, and to detect the conformational changes of helicases during unwinding. Furthermore, these techniques also provide the capability to distinguish helicase heterogeneity and monitor helicase motion at nanometer spatial and millisecond temporal resolutions, ultimately providing new insights into the mechanisms that could not be resolved by ensemble assays. This review outlines the single-molecule techniques that have been utilized for measurements of helicase activities and discusses helicase mechanisms with a focus on functional and mechanistic insights revealed through single-molecule investigations in the past five years.
DOI
Helicases are a diverse group of molecular motors that utilize energy derived from the hydrolysis of nucleoside triphosphates (NTPs) to unwind and translocate along nucleic acids. These enzymes play critical roles in nearly all aspects of nucleic acid metabolism, and consequently, a detailed understanding of helicase mechanisms at the molecular level is essential. Over the past few decades, single-molecule techniques, such as optical tweezers, magnetic tweezers, laminar flow, fluorescence resonance energy transfer (FRET), and DNA curtains, have proved to be powerful tools to investigate the functional properties of both DNA and RNA helicases. These approaches allow researchers to manipulate single helicase molecules, perturb their free energy landscape to probe the chemo-mechanical activities of these motors, and to detect the conformational changes of helicases during unwinding. Furthermore, these techniques also provide the capability to distinguish helicase heterogeneity and monitor helicase motion at nanometer spatial and millisecond temporal resolutions, ultimately providing new insights into the mechanisms that could not be resolved by ensemble assays. This review outlines the single-molecule techniques that have been utilized for measurements of helicase activities and discusses helicase mechanisms with a focus on functional and mechanistic insights revealed through single-molecule investigations in the past five years.
DOI
An electrodynamics-Langevin dynamics (ED-LD) approach to simulate metal nanoparticle interactions and motion
N. Sule, S. A. Rice, S. K. Gray, and N. F. Scherer
Understanding the formation of electrodynamically interacting assemblies of metal nanoparticles requires accurate computational methods for determining the forces and propagating trajectories. However, since computation of electromagnetic forces occurs on attosecond to femtosecond timescales, simulating the motion of colloidal nanoparticles on milliseconds to seconds timescales is a challenging multi-scale computational problem. Here, we present a computational technique for performing accurate simulations of laser-illuminated metal nanoparticles. In the simulation, we self-consistently combine the finite-difference time-domain method for electrodynamics (ED) with Langevin dynamics (LD) for the particle motions. We demonstrate the ED-LD method by calculating the 3D trajectories of a single 100-nm-diameter Ag nanoparticle and optical trapping and optical binding of two and three 150-nm-diameter Ag nanoparticles in simulated optical tweezers. We show that surface charge on the colloidal metal nanoparticles plays an important role in their optically driven self-organization. In fact, these simulations provide a more complete understanding of the assembly of different structures of two and three Ag nanoparticles that have been observed experimentally, demonstrating that the ED-LD method will be a very useful tool for understanding the self-organization of optical matter.
DOI
Understanding the formation of electrodynamically interacting assemblies of metal nanoparticles requires accurate computational methods for determining the forces and propagating trajectories. However, since computation of electromagnetic forces occurs on attosecond to femtosecond timescales, simulating the motion of colloidal nanoparticles on milliseconds to seconds timescales is a challenging multi-scale computational problem. Here, we present a computational technique for performing accurate simulations of laser-illuminated metal nanoparticles. In the simulation, we self-consistently combine the finite-difference time-domain method for electrodynamics (ED) with Langevin dynamics (LD) for the particle motions. We demonstrate the ED-LD method by calculating the 3D trajectories of a single 100-nm-diameter Ag nanoparticle and optical trapping and optical binding of two and three 150-nm-diameter Ag nanoparticles in simulated optical tweezers. We show that surface charge on the colloidal metal nanoparticles plays an important role in their optically driven self-organization. In fact, these simulations provide a more complete understanding of the assembly of different structures of two and three Ag nanoparticles that have been observed experimentally, demonstrating that the ED-LD method will be a very useful tool for understanding the self-organization of optical matter.
DOI
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