High efficiency penetration of antibody-immobilized nanoneedle thorough plasma membrane for in situ detection of cytoskeletal proteins in living cells.

BACKGROUND: The field of structural dynamics of cytoskeletons in living cells is gathering wide interest, since better understanding of cytoskeleton intracellular organization will provide us with not only insights into basic cell biology but may also enable development of new strategies in regenerative medicine and cancer therapy, fields in which cytoskeleton-dependent dynamics play a pivotal role. The nanoneedle technology is a powerful tool allowing for intracellular investigations, as it can be directly inserted into live cells by penetrating through the plasma membrane causing minimal damage to cells, under the precise manipulation using atomic force microscope. Modifications of the nanoneedles using antibodies have allowed for accurate mechanical detection of various cytoskeletal components, including actin, microtubules and intermediate filaments. However, successful penetration of the nanoneedle through the plasma membrane has been shown to vary greatly between different cell types and conditions. In an effort to overcome this problem and improve the success rate of nanoneedle insertion into the live cells, we have focused here on the fluidity of the membrane lipid bilayer, which may hinder nanoneedle penetration into the cytosolic environment. RESULTS: We aimed to reduce apparent fluidity of the membrane by either increasing the approach velocity or reducing experimental temperatures. Although changes in approach velocity did not have much effect, lowering the temperature was found to greatly improve the detection of unbinding forces, suggesting that alteration in the plasma membrane fluidity led to increase in nanoneedle penetration. CONCLUSIONS: Operation at a lower temperature of 4 °C greatly improved the success rate of nanoneedle insertion to live cells at an optimized approach velocity, while it did not affect the binding of antibodies immobilized on the nanoneedle to vimentins for mechanical detection. As these experimental parameters can be applied to various cell types, these results may improve the versatility of the nanoneedle technology to other cell lines and platforms.

Correlated multiphoton holes: absence of multiphoton coincidence events.

We generate bipartite states of light which exhibit an absence of multiphoton coincidence events between two modes amid a constant background flux. These "correlated photon holes" are produced by mixing a coherent state and relatively weak spontaneous parametric down-conversion by using a balanced beam splitter. Correlated holes with arbitrarily high photon numbers may be obtained by adjusting the relative phase and amplitude of the inputs. We measure states of up to five photons and verify their nonclassicality. The scheme provides a route for observation of high-photon-number nonclassical correlations without requiring intense quantum resources.

Classical bound for Mach-Zehnder superresolution.

The employment of path-entangled multiphoton states enables measurement of phase with enhanced precision. It is common practice to demonstrate the unique properties of such quantum states by measuring superresolving oscillations in the coincidence rate of a Mach-Zehnder interferometer. Similar oscillations, however, have also been demonstrated in various configurations using classical light only; making it unclear what, if any, are the classical limits of this phenomenon. Here we derive a classical bound for the visibility of superresolving oscillations in a Mach-Zehnder interferometer. This provides an easy to apply, fundamental test of nonclassicality. We apply this test to experimental multiphoton coincidence measurements obtained using photon number resolving detectors. Mach-Zehnder superresolution is found to be a highly distinctive quantum effect.

Observation of a localization transition in quasiperiodic photonic lattices.

We report the observation of the signature of a localization phase transition for light in one-dimensional quasiperiodic photonic lattices, by directly measuring wave transport inside the lattice. Below the predicted transition point an initially narrow wave packet expands as it propagates, while above the transition expansion is fully suppressed. In addition, we measure the effect of focusing nonlinear interaction on the propagation and find it increases the width of the localized wave packets.

Effect of nonlinearity on adiabatic evolution of light.

We investigate the effect of nonlinearity in a system described by an adiabatically evolving Hamiltonian. Experiments are conducted in a three-core waveguide structure that is adiabatically varying with distance, in analogy to the stimulated Raman adiabatic passage process in atomic physics. In the linear regime, the system exhibits an adiabatic power transfer between two waveguides which are not directly coupled, with negligible power recorded in the intermediate coupling waveguide. In the presence of nonlinearity the adiabatic light passage is found to critically depend on the excitation power. We show how this effect is related to the destruction of the dark state formed in this configuration.

Femtosecond pulse shaping using a two-dimensional liquid-crystal spatial light modulator.

We introduce a programmable, high-rate scanning femtosecond pulse shaper based on a two-dimensional liquid crystal on a silicon spatial light modulator (SLM). While horizontal resolution of 1920 addressable pixels provides superior fidelity for generating complex waveforms, scanning across the vertical dimension (1080 pixels) has been used to facilitate at least 3 orders of magnitude speed increase as compared with typical liquid-crystal SLM-based pulse shapers. An update rate in excess of 100 kHz is demonstrated.

Dentin micro-architecture using harmonic generation microscopy.

OBJECTIVES: We present a novel way to create high-resolution three-dimensional images of tooth dentin by harmonic generation scanning laser microscopy. METHODS: The images were taken using a pulsed infrared laser. Three-dimensional reconstruction enables the visualization of individual tubules and the collagen fibrils mesh around them with an optical resolution of approximately 1 microm. RESULTS: The images show micro-morphological details of the dentinal tubules as well as the collagen fibrils at a depth of up to about 200 microm. The data show that while collagen fibrils are organized in planes perpendicular to the tubules, close to the dentin enamel junction they lie also along the long axis of the tubules. CONCLUSIONS: The unique 3D information opens the opportunity to study the collagen fibril arrangement in relation to the tubule orientation within the dentin matrix, and may be applied to study the micro-morphology of normal versus altered dentin.

Transformation from an ultrashort pulse to a spatiotemporal speckle by a thin scattering surface.

We have theoretically and experimentally studied the local temporal and spectral characteristics of an ultrashort pulse passing a simple thin diffusive element. We show that as one moves away from the diffuser the pulse evolves into a spatiotemporal speckle.

Incoherent blocker soliton interactions in Kerr waveguide arrays.

We have observed the incoherent interaction between a highly confined (blocker) soliton and wide, moving signal beams of a different wavelength in a one-dimensional discrete Kerr medium. Digital switching of the blocker solitons to successive adjacent channels was measured with increasing signal power via both one and two cascaded interactions in an AlGaAs waveguide array, operations equivalent to a reconfigurable three-output router.

Femtosecond pulse-shape modulation at nanosecond rates.

We report high-rate, computer-controlled femtosecond pulse shaping by use of an electro-optical gallium arsenide optical phased-array modulator with 2304 controlled waveguides. It provides fast modulation speed of both spectral phases and amplitudes. Limited by the driving electronics of our current setup, we were able to update a pulse shape in approximately 30 ns. This technique paves the way toward individual shaping of every single pulse of typical femtosecond mode-locked oscillators.

Nonlinearly induced relaxation to the ground state in a two-level system.

We report the experimental demonstration of a nonlinear process in a two-level system, in which the amplitude of the excited state decays, transferring irreversibly a large fraction of its energy to the ground state, while shedding a part of it into radiation states. The experiments where preformed in a nonlinear optical waveguide, supporting two or three modes. The process is general, and is expected to occur in other nonlinear few level systems such as nonlinear quantum wells and Bose-Einstein condensates.

Beam interactions with a blocker soliton in one-dimensional arrays.

We investigate experimentally and numerically the interaction of a highly localized, single-channel discrete soliton (blocker) with a wide, tilted beam in a one-dimensional AlGaAs array. In agreement with theory the blocker is observed to discretely shift its position by multiple channels, depending on the intensity and relative phase of the tilted beam.

Nonlinear beam interactions in 1D discrete Kerr systems.

The interaction between parallel beams in one-dimensional discrete Kerr systems has been investigated using arrays of coupled channel waveguides. The experiments were performed in AlGaAs waveguides at 1550 nm which corresponds to photon energies just below one half the semiconductor's bandgap. The input intensity and relative input phase between the input beams was varied and the output intensity patterns were recorded. Observed was behavior ranging from a linear response, to soliton interactions between moderately and then strongly localized spatial solitons. Finally the influence of multiphoton absorption and asymmetric beam inputs on these interactions was investigated at very high intensities.

Nonlinear optical beam interactions in waveguide arrays.

We report our investigation of Kerr nonlinear beam interactions in discrete systems. The influence of power and the relative phase between two Gaussian shaped beams was investigated in detail by performing numerical simulations of the discrete nonlinear Schrödinger equation and comparing the results with experiments done in AlGaAs waveguide arrays. Good agreement between theory and experiment was obtained.

Experimental observation of discrete modulational instability.

We report the first experimental observation of modulation instability in a discrete optical nonlinear array.

Femtosecond pulse-shape modulation at kilohertz rates.

We demonstrate a new scanning femtosecond pulse-shaping technique that allows pulse shapes to be modulated at kilohertz rates. This technique is particularly useful for lock-in measurements in which the signal is synchronized with the alternating pulse shapes. The pulse-shape lock-in technique is demonstrated in resonant coherent anti-Stokes Raman scattering, where it is shown to significantly improve the ratio of the resonant signal to both the nonresonant background and to noise.

Gap solitons in waveguide arrays.

Bright and dark spatial gap solitons are demonstrated in waveguide arrays. These gap solitons travel across the array at zero transverse velocity, in complete analogy with stationary (immobile) temporal gap solitons. Furthermore, the launching configuration for observing these stationary gap solitons is shown to be the analog of an "ideal experiment" for observing stationary temporal gap solitons, never observed so far. A clear distinction is established between the family of Floquet-Bloch solitons in general and discrete solitons in particular, and the limiting case of gap solitons.

Strong spatiotemporal localization in a silica nonlinear waveguide array.

We investigate the propagation of short, intense laser pulses in arrays of coupled silica waveguides, in the anomalous dispersion regime. The nonlinearity induces trapping of the pulse in a single waveguide, over a wide range of input parameters. A sharp transition is observed for single waveguide excitation, from strong diffraction at low powers to strong localization at high powers.

Discrete vector solitons in Kerr nonlinear waveguide arrays.

We report the first experimental observation of discrete vector solitons in AlGaAs nonlinear waveguide arrays. These self-trapped states are possible through the coexistence of two orthogonally polarized fields and are stable in spite of the presence of four-wave mixing effects. We demonstrate that at sufficiently high power levels the two polarizations lock into a highly localized vector discrete soliton that would have been otherwise impossible in the absence of either one of these two components.

Observation of mutually trapped multiband optical breathers in waveguide arrays.

Multiphoton fluorescence is used for the direct observation of a new class of breathers in waveguide arrays, which are a coherent superposition of Floquet-Bloch solitons of different bands. These Floquet-Bloch breathers oscillate along their spatial propagation axis, and possess several novel properties. Some behavior of these breathers is readily understood intuitively in terms of the band structure of the waveguide array and the properties of discrete solitons.