Full-field optical sectioning and three-dimensional localization of fluorescent particles using focal plane modulation

We present a technique for imaging fluorescent particles based on the axial modulation of the objective's focal plane position. This technique provides full-field optical sectioning and can be used to localize the fluorophores in three dimensions. We describe the technique and apply it to image 200 nm diameter fluorescent beads immobilized in a gel. We show that full-field optical sectioning is obtained and that the beads are localized with a precision of 10 nm in the transverse plane and 14 nm in the axial direction.     

Three-dimensional tracking of small spheres in focused laser beams: influence of the detection angular aperture

Back-focal-plane interferometry is a method capable of determining the three-dimensional position of a particle with high precision (< 3 nm) at high sampling rates (1 MHz). We investigated theoretically the performance of such a system for dielectric spheres with diameters D = 0.53-3 microm and for metallic spheres with D < or = 300 nm. Good sensitivity and linearity were achieved for a detection angular aperture sin(alpha) of no more than 0.5. A value of sin(alpha) > 0.7 should be used only for dielectric spheres with diameters approximately equal to the laser wavelength. Harmonic optical traps can be calibrated by measurement of the thermal motion of the sphere. We performed Brownian dynamics simulations and subsequent thermal noise analyses to prove that the wrong sin(alpha) incorrectly suggests an increased and nonharmonic axial trapping potential.     

Trapping and manipulation of low-index particles in a two-dimensional interferometric optical trap

We demonstrate optical trapping and manipulation of low-index spheres in two dimensions, using the pattern produced by two interfering plane waves. This technique shows, for what is believed to be the first time, alignment of an array of hollow spheres and simultaneous manipulation of high- and low-index particles in the horizontal plane. Furthermore, rodlike particles (up to 30microm in length) are manipulated simultaneously with the low-index particles. This technique offers a practical method for manipulating bubbles, low-index droplets, or rodlike biological samples.     

光学微操纵过程的轴平面显微成像技术

光学俘获技术利用光与物质相互作用产生的光势阱效应来实现对微粒的操控,已经成功应用于生物医学、材料科学等交叉领域.在对微粒进行三维俘获时,传统的宽场光学显微技术只能观测到某一平面内微粒的横向运动,对微粒沿轴向运动的观测受到很大限制.本文将轴平面显微成像技术引入光学微粒操控研究中,利用45?倾斜的反射镜把微粒的轴向运动信息转换到横向平面进行观测,与传统宽场显微成像技术相结合,实现了对二氧化硅小球俘获过程横向和轴向运动的同步观测.该成像方法无需扫描和数据重构,具有实时快速等优点,在新型光束光镊、厚样品三维观测和成像等领域具有潜在的应用价值.     

Real-time three-dimensional optical micromanipulation of multiple particles and living cells

Counterpropagating light fields provide a stationary optical potential well for a Brownian particle. Introducing variability in the relative strengths of the counterpropagating beams allows us to create a more general configuration-the optical elevator. An optical elevator dynamically controls the axial location of the potential minimum where the particle finds a stable equilibrium position. We describe the implementation of multiple real-time reconfigurable optical elevators with the generalized phase contrast method for dynamic manipulation of polystyrene spheres and yeast cells S. cerevisiae in three dimensions.     

Three-dimensional holographic fluorescence microscopy

Most commonly used methods for three-dimensional (3D) fluorescence microscopy make use of sectioning techniques that require that the object be physically scanned in a series of two-dimensional (2D) sections along the z axis. The main drawback in these approaches is the need for these sequential 2D scans. An alternative approach to fluorescence imaging in three dimensions has been developed that is based on optical scanning holography. This novel approach requires only a 2D scan to record 3D information. Holograms of 15-microm fluorescent latex beads with longitinal separation of ~2 mm have been recorded and reconstructed. To our knowledge, this is the first time holograms of fluorescent specimens have been recorded by an optical holographic technique.     

Active Brownian motion tunable by light

Active Brownian particles are capable of taking up energy from their environment and converting it into directed motion; examples range from chemotactic cells and bacteria to artificial micro-swimmers. We have recently demonstrated that Janus particles, i.e.?gold-capped colloidal spheres, suspended in a critical binary liquid mixture perform active Brownian motion when illuminated by light. In this paper, we investigate in more detail their swimming mechanism, leading to active Brownian motion. We show that the illumination-borne heating induces a local asymmetric demixing of the binary mixture, generating a spatial chemical concentration gradient which is responsible for the particle's self-diffusiophoretic motion. We study this effect as a function of the functionalization of the gold cap, the particle size and the illumination intensity: the functionalization determines what component of the binary mixture is preferentially adsorbed at the cap and the swimming direction (towards or away from the cap); the particle size determines the rotational diffusion and, therefore, the random reorientation of the particle; and the intensity tunes the strength of the heating and, therefore, of the motion. Finally, we harness this dependence of the swimming strength on the illumination intensity to investigate the behavior of a micro-swimmer in a spatial light gradient, where its swimming properties are space-dependent.     

Talbot holographic illumination nonscanning (THIN) fluorescence microscopy

Optical sectioning techniques offer the ability to acquire three‐dimensional information from various organ tissues by discriminating between the desired in‐focus and out‐of‐focus (background) signals. Alternative techniques to confocal, such as active structured illumination, exist for fast optically sectioned images, but they require individual axial planes to be imaged consecutively. In this article, an imaging technique (THIN), by utilizing active Talbot illumination in 3D and multiplexed holographic Bragg filters for depth discrimination, is demonstrated for imaging in vivo 3D biopsy without mechanical or optical axial scanning.     

Diffusive motion and recurrence on an idealized Galton board

We study a mechanical model known as a Galton board--a particle rolling on a tilted plane under gravitation and bouncing off a periodic array of rigid pegs. Incidentally, this model is identical to a periodic Lorentz gas where an electron is driven by a uniform electric field. Previous heuristic and experimental studies have suggested that the particle's speed v(t) should grow as t(1/3) and its coordinate x(t) as t(2/3). We find exact limit distributions for the rescaled velocity t(-1/3)v(t) and position t(-2/3)x(t). In addition, we determine that the particle's motion is recurrent; i.e., the particle comes back to the top of the board with a probability of one.     

Numerical Simulation of Random Close Packings in Particle Deformation from Spheres to Cubes

Variation of packing density in particle deforming from spheres to cubes is studied. A new model is presented to describe particle deformation between different particle shapes. Deformation is simulated by relative motion of component spheres in the sphere assembly model of a particle. Random close packings of particles in deformation form spheres to cubes are simulated with an improved relaxation algorithm. Packings in both 2D and 3D cases are simulated. With the simulations, we find that the packing density increases while the particle sphericity decreases in the deformation. Spheres and cubes give the minimum (0.6404) and maximum (0.7755) of packing density in the deformation respectively. In each deforming step, packings starting from a random configuration and from the final packing of last deforming step are both simulated. The packing density in the latter case is larger than the former in two dimensions, but is smaller in three dimensions. The deformation model can be applied to other particle shapes as well.     

Human cervical carcinoma detection and glucose monitoring in blood micro vasculatures with swept source OCT

We report a pilot method, i.e., speckle variance (SV) and structured optical coherence tomography to visualize normal and malignant blood microvasculature in three and two dimensions and to monitor the glucose levels in blood by analyzing the Brownian motion of the red blood cells. The technique was applied on nude live mouse’s skin and the obtained images depict the enhanced intravasculature network forum up to the depth of ～2 mm with axial resolution of ～8 μm. Microscopic images have also been obtained for both types of blood vessels to observe the tumor spatially. Our SV-OCT methodologies and results give satisfactory techniques in real time imaging and can potentially be applied during therapeutic techniques such as photodynamic therapy as well as to quantify the higher glucose levels injected intravenously to animal by determining the translation diffusion coefficient.     

Rotational motion in sensorless freehand three-dimensional ultrasound

Freehand three-dimensional ultrasound is usually acquired with a position sensor attached to the ultrasound probe. However, position sensors can be expensive, obtrusive and difficult to calibrate. For this reason, there has been much research on alternative, image-based techniques, with in-plane motion tracked using conventional image registration methods, and out-of-plane motion inferred from the decorrelation between nearby B-scans. However, since out-of-plane motion is not the only source of decorrelation, image-based positions determined in this way suffer from cumulative drift errors. In this paper, we consider the effect of probe rotation on correlation and how this affects the position estimates. We then present a novel technique to compensate for out-of-plane rotations, by making use of orientation measurements from an unobtrusive sensor. Using simulations and in vitro experiments, we demonstrate that the technique is able to reduce the drift error in elevational positioning by 57% on average.     

Motion of nonspherical dust particle under the action of electromagnetic radiation

Equation of motion of realistically shaped particle in the circumstellar dust shell is derived under the action of electromagnetic radiation including the gravity of central body. The effect is considered to the accuracy , where is particle's velocity in a given inertial frame of reference and c is the speed of light. Equation of motion is expressed in terms of particle's optical properties, standardly used in optics for stationary particles.

Application to nonspherical dust particle in the Solar System with initial orbital elements identical to those of comet Encke is presented as an example. It is shown that the motion of nonspherical submicron- and small micron-sized particle may significantly differ from the motion for spherical particle of an identical volume.     

Optical levitation of absorbing particles with a nominally Gaussian laser beam

We use a Gaussian laser beam to study the levitation of absorbing Mie particles. Several metal oxide particles are stably levitated, and their movement over time is recorded. Our studies show that the position of each particle is highly dependent on the other particles' locations. The observations are explained by the phenomenon of thermal creep. The increased local pressure that is due to a temperature gradient along the particle's surface induces levitation. The particles rest close to minima in the intensity distribution near the optical axis. An experiment is suggested that can be used to locate these minima in a laser beam.     

Real-time flow imaging by removing texture pattern artifacts in spectral-domain optical Doppler tomography

We present a new, simple method to suppress texture pattern artifacts induced by the optical heterogeneity of tissues to improve the performance of flow imaging for real-time phase-resolved optical Doppler tomography. The method performs transverse scanning of the probe beam in the forward and then reverse directions, and it takes average of the spatial phase changes between them to obtain the final velocity image. It relies on the fact that the phase changes between successive axial scans due to the optical heterogeneity of the sample are time independent, while those due to the moving particles are time dependent. We experimentally demonstrate this method by real-time imaging of a flow phantom.     

3D homogeneity study in PMMA layers using a Fourier domain OCT system

Micro-metallic particles embedded in polymers are now widely used in several industrial applications in order to modify the mechanical properties of the bulk. A uniform distribution of these particles inside the polymers is highly desired for instance, when a biological backscattering is simulated or a bio-framework is designed. A 3D Fourier domain optical coherence tomography system to detect the polymer's internal homogeneity is proposed. This optical system has a 2D camera sensor array that records a fringe pattern used to reconstruct with a single shot the tomographic image of the sample. The system gathers the full 3D tomographic and optical phase information during a controlled deformation by means of a motion linear stage. This stage avoids the use of expensive tilting stages, which in addition are commonly controlled by piezo drivers. As proof of principle, a series of different deformations were proposed to detect the uniform or non-uniform internal deposition of copper micro particles. The results are presented as images coming from the 3D tomographic micro reconstruction of the samples, and the 3D optical phase information that identifies the in-homogeneity regions within the Poly methyl methacrylate (PMMA) volume.     

On classical electrodynamics of point particles and mass renormalization: Some preliminary results

Apparently, no rigorous results exist for the dynamics of a classical point particle interacting with the electromagnetic field, as described by the standard Maxwell-Lorentz equations. Some results are given here for the corresponding linearized system (dipole approximation) in the presence of a mechanical linear restoring force. We consider a regularization of the system (Pauli-Fierz model), and explicitly solve the Cauchy problem in terms of normal modes. Then we study the limit of the particle's motion as the regularization is removed. We prove that the particle's motion corresponding to smooth initial data for the field has a well-defined limit if mass is renormalized, while the motion is trivial (i.e. the particle does not move at all) if mass is not renormalized. Moreover, the limit particle's motion corresponding to an interesting class of initial data satifies exactly the Abraham-Lorentz-Dirac equation. Finally, for generic initial data the limit motion is runaway.     

Localizing and Characterizing Colloidal Particles Scattering Using Lens-free Holographic Microscopy

Lens-free holographic microscopy could achieve both improved resolution and field of view(FOV), which has huge potential applications in biomedicine, fluid mechanics and soft matter physics. Unfortunately, due to the limited sensor pixel size, target objects could not be located to a satisfactory level. Recent studies have shown that electromagnetic scattering can be fitted to digital holograms to obtain the 3 D positions of isolated colloidal spheres with nanometer precision and millisecond temporal resolution. Here, we describe a lens-free holographic imaging technique that fits multi-sphere superposition scattering to digital holograms to obtain in situ particle position and model parameters: size and refractive index of colloidal spheres. We show that the proposed method can be utilized to analyze the location and character of colloidal particles under large FOV with high density.     

Quantitative analysis of depolarization of backscattered light by stochastically inhomogeneous dielectric particles

We determine the relationship between the depolarization properties of inhomogeneous particles and the statistical parameters of their internal refractive-index distributions. Our analysis demonstrates that the linear depolarization ratio of backscattered light by an inhomogeneous particle is approximately proportional to both the squared standard deviation and the squared correlation length of the particle's internal refractive-index distribution. We verify this result by conducting rigorous numerical studies using the finite-difference time-domain method. This improved understanding of light depolarization by inhomogeneous structures may enhance polarization-based biomedical optical imaging techniques.     

轴向多光阱微粒捕获与实时直接观测技术

传统的光镊技术使用单个物镜同时进行光学捕获与显微成像,使得捕获与成像区域被限制在物镜焦平面附近,无法同时观察到沿光轴方向(即Z向)捕获的多个微粒.本文提出一种轴平面(XZ平面)GerchbergSaxton迭代算法来产生沿轴向分布的多光阱阵列,将轴平面成像技术与光镊结合,实现了沿轴向对二氧化硅微球的多光阱同时捕获与实时观测.通过视频分析法测量了多个二氧化硅微球在轴向光镊阵列中的布朗运动,并标定了光阱刚度.本文提出的轴向多光阱微粒捕获与实时观测技术为光学微操纵提供了一个新的观测视角和操纵方法,为生物医学、物理学等相关领域研究提供了一种新的技术手段.