Abstract: Optical-domain subsampling enables Fourier-domain OCT imaging at high-speeds and extended depth ranges while limiting the required acquisition bandwidth. To perform optical-domain subsampling, a wavelength-stepped rather than a wavelength-swept source is required. This preliminary study introduces a novel design for a rapid wavelength-stepped laser source that uses dispersive fibers in combination with a fast lithium-niobate modulator to achieve wavelength selection. A laser with 200 GHz wavelength-stepping and a sweep rate of 9 MHz over a 94 nm range at a center wavelength of 1550 nm is demonstrated. A reconfiguration of this source design to a continuous wavelength-swept light for conventional Fourier-domain OCT is also demonstrated.
(140.3600) Lasers, tunable; (110.4500) Optical coherence tomography
Schemes for X-ray imaging single protein molecules using new x-ray sources, like x-ray free electron lasers (XFELs), require processing many frames of data that are obtained by taking temporally short snapshots of identical molecules, each with a random and unknown orientation. Due to the small size of the molecules and short exposure times, average signal levels of much less than 1 photon/pixel/frame are expected, much too low to be processed using standard methods. One approach to process the data is to use statistical methods developed in the EMC algorithm (Loh & Elser, Phys. Rev. E, 2009) which processes the data set as a whole. In this paper we apply this method to a real-space tomographic reconstruction using sparse frames of data (below 10−2 photons/pixel/frame) obtained by performing x-ray transmission measurements of a low-contrast, randomly-oriented object. This extends the work by Philipp et al. (Optics Express, 2012) to three dimensions and is one step closer to the single molecule reconstruction problem.
(000.2190) Experimental physics; (040.7480) X-rays, soft x-rays, extreme ultraviolet (EUV); (100.6950) Tomographic image processing; (110.4155) Multiframe image processing; (110.4280) Noise in imaging systems; (110.6955) Tomographic imaging; (110.7440) X-ray imaging; (340.7440) X-ray imaging
Oblique back-illumination microscopy (OBM) provides high resolution, sub-surface phase-gradient images from arbitrarily thick samples. We present an image formation theory for OBM and demonstrate that OBM lends itself to volumetric imaging because of its capacity for optical sectioning. In particular, OBM can provide extended depth of field (EDOF) images from single exposures, by rapidly scanning the focal plane with an electrically tunable lens. These EDOF images can be further enhanced by deconvolution. We corroborate our theory with experimental volumetric images obtained from transparent bead samples and mouse cortical brain slices.
(110.0180) Microscopy; (350.5030) Phase; (120.5050) Phase measurement
A new computational imaging technique, termed Fourier ptychographic microscopy (FPM), uses a sequence of low-resolution images captured under varied illumination to iteratively converge upon a high-resolution complex sample estimate. Here, we propose a mathematical model of FPM that explicitly connects its operation to conventional ptychography, a common procedure applied to electron and X-ray diffractive imaging. Our mathematical framework demonstrates that under ideal illumination conditions, conventional ptychography and FPM both produce datasets that are mathematically linked by a linear transformation. We hope this finding encourages the future cross-pollination of ideas between two otherwise unconnected experimental imaging procedures. In addition, the coherence state of the illumination source used by each imaging platform is critical to successful operation, yet currently not well understood. We apply our mathematical framework to demonstrate that partial coherence uniquely alters both conventional ptychography’s and FPM’s captured data, but up to a certain threshold can still lead to accurate resolution-enhanced imaging through appropriate computational post-processing. We verify this theoretical finding through simulation and experiment.
(110.1758) Computational imaging; (110.4980) Partial coherence in imaging; (070.7425) Quasi-probability distribution functions; (080.5084) Phase space methods of analysis
Rotating mirror cameras represent a workhorse technology for high speed imaging in the MHz framing regime. The technique requires that the target image be swept across a series of juxtaposed CCD sensors, via reflection from a rapidly rotating mirror. Employing multiple sensors in this fashion can lead to spatial jitter in the resultant video file, due to component misalignments along the individual optical paths to each CCD. Here, we highlight that static and dynamic fiducials can be exploited as an effective software-borne countermeasure to jitter, suppressing the standard deviation of the corrected file relative to the raw data by up to 88.5% maximally, and 66.5% on average over the available range of framing rates. Direct comparison with industry-standard algorithms demonstrated that our fiducial-based strategy is as effective at jitter reduction, but typically also leads to an aesthetically superior final form in the post-processed video files.
We report size-based sorting of micro- and sub-micron particles using optical forces on a planar optofluidic chip. Two different combinations of fluid flow and optical beam directions in liquid-core waveguides are demonstrated. These methods allow for tunability of size selection and sorting with efficiencies as high as 100%. Very good agreement between experimental results and calculated particle trajectories in the presence of flow and optical forces is found.
(130.3120) Integrated optics devices; (230.7390) Waveguides, planar; (350.4855) Optical tweezers or optical manipulation
We experimentally investigated the nonlinear optical interaction between the instantaneous four-wave mixing and the cascaded quadratic frequency conversion in commonly used nonlinear optical KTP and LiNbO3 with the aim of a possible background suppression of the non-resonant background in coherent anti-Stokes Raman scattering. The possibility of background-free heterodyne coherent anti-Stokes Raman scattering microspectroscopy is investigated at the interface formed by a liquid (isopropyl alcohol) and a nonlinear crystal (LiNbO3).
(190.7110) Ultrafast nonlinear optics; (190.4720) Optical nonlinearities of condensed matter; (290.5860) Scattering, Raman; (180.5655) Raman microscopy
Regeneration of the intestinal epithelium after injury or during pathogenesis is a dynamic cellular process critical for host immunity. However, current epithelial injury models provide poor spatial control, complicating the study of precise cellular responses. Here we developed endoscopic femtosecond-laser surgery capable of generating acute tissue injury. A side-view probe provides a convenient access to the distal colon in the mouse in vivo and allows real-time intraoperative monitoring as well as pre- and post-surgery examinations via multiphoton imaging. The photo-induced damage showed a nonlinear dependence on laser intensity. At an optical power of 200 mW (2.5 nJ per pulse), scanning the beam focus over 300x300 µm2 area in the colonic mucosa generated substantial vascular damages within 30 s. We confirmed the localized tissue damage and the physiologic regeneration of the disrupted epithelium by in situ barrier function assays, validating the animal model for epithelial regeneration following injury. The femtosecond endosurgery technique is applicable to various experimental models based on laser-induced perturbations.
(170.2150) Endoscopic imaging; (180.4315) Nonlinear microscopy; (170.1020) Ablation of tissue; (000.1430) Biology and medicine
Owing to its superior resolution, intravascular optical coherence tomography (IVOCT) is a promising tool for imaging the microstructure of coronary artery walls. However, IVOCT does not identify chemicals and molecules in the tissue, which is required for a more complete understanding and accurate diagnosis of coronary disease. Here we present a dual-modality imaging system and catheter that uniquely combines IVOCT with diffuse near-infrared spectroscopy (NIRS) in a single dual-modality imaging device for simultaneous acquisition of microstructural and compositional information. As a proof-of-concept demonstration, the device has been used to visualize co-incident microstructural and spectroscopic information obtained from a diseased cadaver human coronary artery.
(110.4500) Optical coherence tomography; (170.6510) Spectroscopy, tissue diagnostics; (110.0113) Imaging through turbid media
An adaptive optics scanning laser ophthalmoscope (AO-SLO) is adapted to provide optical coherence tomography (OCT) imaging. The AO-SLO function is unchanged. The system uses the same light source, scanning optics, and adaptive optics in both imaging modes. The result is a dual-modal system that can acquire retinal images in both en face and cross-section planes at the single cell level. A new spectral shaping method is developed to reduce the large sidelobes in the coherence profile of the OCT imaging when a non-ideal source is used with a minimal introduction of noise. The technique uses a combination of two existing digital techniques. The thickness and position of the traditionally named inner segment/outer segment junction are measured from individual photoreceptors. In-vivo images of healthy and diseased human retinas are demonstrated.
(110.4500) Optical coherence tomography; (110.1080) Active or adaptive optics; (170.4460) Ophthalmic optics and devices; (170.4470) Ophthalmology
Localization-based superresolution imaging is dependent on finding the positions of individualfluorophores in a sample by fitting the observed single-molecule intensity pattern to the microscopepoint spread function (PSF). For three-dimensional imaging, system-specific aberrations of theoptical system can lead to inaccurate localizations when the PSF model does not account for theseaberrations. Here we describe the use of phase-retrieved pupil functions to generate a more accuratePSF and therefore more accurate 3D localizations. The complex-valued pupil function containsinformation about the system-specific aberrations and can thus be used to generate the PSF forarbitrary defocus. Further, it can be modified to include depth dependent aberrations. We describethe phase retrieval process, the method for including depth dependent aberrations, and a fastfitting algorithm using graphics processing units. The superior localization accuracy of the pupilfunction generated PSF is demonstrated with dual focal plane 3D superresolution imaging ofbiological structures.
(100.6640) Superresolution; (180.2520) Fluorescence microscopy; (100.5070) Phase retrieval; (100.6890) Three-dimensional image processing
In super-resolution imaging techniques based on single-molecule switching and localization, the time to acquire a super-resolution image is limited by the maximum density of fluorescent emitters that can be accurately localized per imaging frame. In order to increase the imaging rate, several methods have been recently developed to analyze images with higher emitter densities. One powerful approach uses methods based on compressed sensing to increase the analyzable emitter density per imaging frame by several-fold compared to other reported approaches. However, the computational cost of this approach, which uses interior point methods, is high, and analysis of a typical 40 µm x 40 µm field-of-view super-resolution movie requires thousands of hours on a high-end desktop personal computer. Here, we demonstrate an alternative compressed-sensing algorithm, L1-Homotopy (L1H), which can generate super-resolution image reconstructions that are essentially identical to those derived using interior point methods in one to two orders of magnitude less time depending on the emitter density. Moreover, for an experimental data set with varying emitter density, L1H analysis is ~300-fold faster than interior point methods. This drastic reduction in computational time should allow the compressed sensing approach to be routinely applied to super-resolution image analysis.
(170.2520) Fluorescence microscopy; (100.6640) Superresolution; (110.2960) Image analysis
Low threshold lasers based on rare-earth elements have enabled numerous scientific discoveries and innovations in industry. However, pushing the threshold into the sub-microwatt regime has been stymied by a fundamental material phenomenon. Specifically, rare earth dopants form clusters which quench emission and reduce efficiency. Here, we fabricate resonant cavity lasers from neodymium-doped silica films containing alumina. The alumina prevents the clustering of the Neodymium, enabling the lasers to achieve thresholds of 530nanoWatts at room temperature.
(130.0130) Integrated optics; (140.3530) Lasers, neodymium; (140.4780) Optical resonators; (160.5690) Rare-earth-doped materials; (160.6060) Solgel
An innovative iterative search method called the synthetic phase-shifting (SPS) algorithm is proposed. This search algorithm is used for maximum-likelihood (ML) estimation of a wavefront that is described by a finite set of Zernike Fringe polynomials. In this paper, we estimate the coefficient, or parameter, values of the wavefront using a single interferogram obtained from a point-diffraction interferometer (PDI). In order to find the estimates, we first calculate the squared-difference between the measured and simulated interferograms. Under certain assumptions, this squared-difference image can be treated as an interferogram showing the phase difference between the true wavefront deviation and simulated wavefront deviation. The wavefront deviation is the difference between the reference and the test wavefronts. We calculate the phase difference using a traditional phase-shifting technique without physical phase-shifters. We present a detailed forward model for the PDI interferogram, including the effect of the finite size of a detector pixel. The algorithm was validated with computational studies and its performance and constraints are discussed. A prototype PDI was built and the algorithm was also experimentally validated. A large wavefront deviation was successfully estimated without using null optics or physical phase-shifters. The experimental result shows that the proposed algorithm has great potential to provide an accurate tool for non-null testing.
(100.2650) Fringe analysis; (110.3175) Interferometric imaging; (120.5050) Phase measurement
Light field microscopy is a new technique for high-speed volumetric imaging of weakly scattering or fluorescent specimens. It employs an array of microlenses to trade off spatial resolution against angular resolution, thereby allowing a 4-D light field to be captured using a single photographic exposure without the need for scanning. The recorded light field can then be used to computationally reconstruct a full volume. In this paper, we present an optical model for light field microscopy based on wave optics, instead of previously reported ray optics models. We also present a 3-D deconvolution method for light field microscopy that is able to reconstruct volumes at higher spatial resolution, and with better optical sectioning, than previously reported. To accomplish this, we take advantage of the dense spatio-angular sampling provided by a microlens array at axial positions away from the native object plane. This dense sampling permits us to decode aliasing present in the light field to reconstruct high-frequency information. We formulate our method as an inverse problem for reconstructing the 3-D volume, which we solve using a GPU-accelerated iterative algorithm. Theoretical limits on the depth-dependent lateral resolution of the reconstructed volumes are derived. We show that these limits are in good agreement with experimental results on a standard USAF 1951 resolution target. Finally, we present 3-D reconstructions of pollen grains that demonstrate the improvements in fidelity made possible by our method.
(180.6900) Three dimensional microscopy; (180.2520) Fluorescence microscopy; (100.1830) Deconvolution; (100.6950) Tomographic image processing; (100.3190) Inverse problems
Grüneisen parameter is a key temperature-dependent physical characteristic responsible for thermoelastic efficiency of materials. We propose a new methodology for accurate measurements of temperature dependence of Grüneisen parameter in optically absorbing solutions. We use two-dimensional optoacoustic (OA) imaging to improve accuracy of measurements. Our approach eliminates contribution of local optical fluence and absorbance. To validate the proposed methodology, we studied temperature dependence of aqueous cupric sulfate solutions in the range from 22 to 4°C. Our results for the most diluted salt perfectly matched known temperature dependence for the Grüneisen parameter of water. We also found that Grüneisen-temperature relationship for cupric sulfate exhibits linear trend with respect to the concentration. In addition to accurate measurements of Grüneisen changes with temperature, the developed technique provides a basis for future high precision OA temperature monitoring in live tissues.
(170.5120) Photoacoustic imaging; (120.6780) Temperature; (120.6810) Thermal effects
Non-invasive injectable cellular therapeutic strategies based on sustained delivery of physiological levels of BMP-2 for spinal fusion are emerging as promising alternatives, which could provide sufficient fusion without the associated surgical risks. However, these injectable therapies are dependent on bone formation occurring only at the specific target region. In this study, we developed and deployed fluorescence gene reporter tomography (FGRT) to provide information on in vivo cell localization and viability. This information is sought to confirm the ideal placement of the materials with respect to the area where early bone reaction is required, ultimately providing three dimensional data about the future fusion. However, because almost all conventional fluorescence gene reporters require visible excitation wavelengths, current in vivo imaging of fluorescent proteins is limited by high tissue absorption and confounding autofluorescence. We previously administered fibroblasts engineered to produce BMP-2, but is difficult to determine 3-D information of placement prior to bone formation. Herein we used the far-red fluorescence gene reporter, IFP1.4 to report the position and viability of fibroblasts and developed 3-D tomography to provide placement information. A custom small animal, far-red fluorescence tomography system integrated into a commercial CT scanner was used to assess IFP1.4 fluorescence and to demark 3-D placement of encapsulated fibroblasts with respect to the vertebrae and early bone formation as assessed from CT. The results from three experiments showed that the placement of the materials within the spine could be detected. This work shows that in vivo fluorescence gene reporter tomography of cell-based gene therapy is feasible and could help guide cell-based therapies in preclinical models.
(110.6960) Tomography; (170.3010) Image reconstruction techniques; (170.6280) Spectroscopy, fluorescence and luminescence
We demonstrate a 3-D scanning micromirror device that combines 2-D beam scanning with focus control in the same device using micro-electro-mechanical-systems (MEMS) technology. 2-D beam scanning is achieved with a biaxial gimbal structure and focus control is obtained with a deformable mirror membrane surface. The micromirror with 800 micrometer diameter is designed to be sufficiently compact and efficient so that it can be incorporated into an endoscopic imaging probe in the future. The design, fabrication and characterization of the device are described in this paper. Using the focus-tracking MEMS scanning mirror, we achieved an optical scanning range of >16 degrees with <40 V actuation voltage at resonance and a tunable focal length between infinity and 25 mm with <100V applied bias.
(170.2150) Endoscopic imaging; (230.0230) Optical devices; (230.4685) Optical microelectromechanical devices
We predict and realize the targeted wavelength conversion from the 1550-nm band of a fs Er:fiber laser to an isolated band inside 370-850 nm, corresponding to a blue-shift of 700-1180 nm. The conversion utilizes resonant dispersive wave generation in widely available optical fibers with good efficiency (~7%). The converted band has a large pulse energy (~1 nJ), high spectral brightness (~1 mW/nm), and broad Gaussian-like spectrum compressible to clean transform-limited ~17 fs pulses. The corresponding coherent fiber sources open up portable applications of optical parametric oscillators and dual-output synchronized ultrafast lasers.
(190.4370) Nonlinear optics, fibers; (060.5295) Photonic crystal fibers; (190.5530) Pulse propagation and temporal solitons
Surgical procedures as a prelude to optical imaging are a rate-limiting step in experimental neuroscience. Towards automation of these procedures, we describe the use of nonlinear optical techniques to create a thinned skull window for transcranial imaging. Metrology by second harmonic generation was used to map the surfaces of the skull and define a cutting path. Plasma-mediated laser ablation was utilized to cut bone. Mice prepared with these techniques were used to image subsurface cortical vasculature and blood flow. The viability of the brain tissue was confirmed via histological analysis and supports the utility of solely optical techniques for osteotomy and potentially other surgical procedures.
(170.0170) Medical optics and biotechnology; (120.0120) Instrumentation, measurement, and metrology; (180.0180) Microscopy
The evanescent wave of the cylindrical vector field is analyzed using the vector angular spectrum of the electromagnetic beam. Comparison between the contributions of the TE and TM terms of both the propagating and the evanescent waves associated with the cylindrical vector field in free space is demonstrated. The physical pictures of the evanescent wave and the propagating wave are well illustrated from the vectorial structure, which provides a new approach to manipulating laser beams by choosing the states of polarization in the cross-section of the field.
(260.5430) Polarization; (260.2110) Electromagnetic optics; (260.0260) Physical optics; (070.7345) Wave propagation; (070.0070) Fourier optics and signal processing; (050.1940) Diffraction
Simultaneous spatial and temporal focusing (SSTF) provides precise control of the pulse front tilt necessary to achieve nonreciprocal writing in glass. The magnitude of the pulse front tilt may be adjusted over several orders of magnitude. Nonreciprocal writing was observed for a larger range of focal depths within the sample using SSTF, and nonreciprocal ablation patterns on the surface were revealed. Further, the lower numerical aperture (0.04 NA) utilized with SSTF increases the rate of writing. This technique allows channels in microfluidic devices to be prototyped an order of magnitude faster than with current methods.
A challenge for nonlinear imaging in living tissue is to maximize the total fluorescent yield from each fluorophore. We investigated the emission rates of three fluorophores – rhodamine B, a red fluorescent protein, and CdSe quantum dots – while manipulating the phase of the laser excitation pulse at the focus. In all cases a transform-limited pulse maximized the total yield to insure the highest signal-to-noise ratio. Further, we find evidence of fluorescence anti-bleaching in quantum dot samples.
With extremely low material absorption and exceptional surface smoothness, silica-based optical resonators can achieve extremely high cavity quality (Q) factors. However, the intrinsic material limitations of silica (e.g., lack of second order nonlinearity) may limit the potential applications of silica-based high Q resonators. Here we report some results in utilizing layer-by-layer self-assembly to functionalize silica microspheres with nonlinear and plasmonic nanomaterials while maintaining Q factors as high as 107. We compare experimentally measured Q factors with theoretical estimates, and find good agreement.
(060.3510) Lasers, fiber; (140.4780) Optical resonators
Coherent detection through two opposing objectives (4Pi configuration) improves the precision of three-dimensional (3D) single-molecule localization substantially along the axial direction, but suffers from instrument complexity and maintenance difficulty. To address these issues, we have realized 4Pi fluorescence detection by sandwiching the sample between the objective and a mirror, and create interference of direct incidence and mirror-reflected signal at the camera with a spatial light modulator. Multifocal imaging using this single-objective mirror interference scheme offers improvement in the axial localization similar to the traditional 4Pi method. We have also devised several PSF engineering schemes to enable 3D localization with a single emitter image, offering better axial precision than normal single-objective localization methods such as astigmatic imaging.
(180.2520) Fluorescence microscopy; (180.3170) Interference microscopy; (110.6880) Three-dimensional image acquisition; (110.1080) Active or adaptive optics; (350.5730) Resolution