Computational refocusing in phase-unstable polarization-sensitive optical coherence tomography.

We present computational refocusing in polarization-sensitive optical coherence tomography (PS-OCT) to improve spatial resolution in the calculated polarimetric parameters and extend the depth-of-field in phase-unstable, fiber-based PS-OCT systems. To achieve this, we successfully adapted short A-line range phase-stability adaptive optics (SHARP), a computational aberration correction technique compatible with phase-unstable systems, into a Stokes-based PS-OCT system with inter-A-line polarization modulation. Together with the spectral binning technique to mitigate system-induced chromatic polarization effects, we show that computational refocusing improves image quality in tissue polarimetry of swine eye anterior segment ex vivo with PS-OCT. The benefits, drawbacks, and potential applications of computational refocusing in anterior segment imaging are discussed.

Endoscopic imaging of white matter fiber tracts using polarization-sensitive optical coherence tomography.

Polarization sensitive optical coherence tomography (PSOCT) has been shown to image and delineate white matter fibers in a label-free manner by revealing optical birefringence within the myelin sheath using a microscope setup. In this proof-of-concept study, we adapt recent advancements in endoscopic PSOCT to perform depth-resolved imaging of white matter structures deep inside intact porcine brain tissue ex-vivo, through a small, rotational fiber probe. The probe geometry is comparable to microelectrodes currently used in neurosurgical interventions. The presented imaging system is mobile, robust, and uses biologically safe levels of optical radiation making it well suited for clinical translation. In neurosurgery, where accuracy is imperative, endoscopic PSOCT through a narrow-gauge fiber probe could provide intra-operative feedback on the location of critical white matter structures.

Spectral- and Polarization-Dependent Scattering of Gold Nanobipyramids for Exogenous Contrast in Optical Coherence Tomography.

Polarization-sensitive optical coherence tomography (PS-OCT) reveals the subsurface microstructure of biological tissue and provides information regarding the polarization state of light backscattered from tissue. Complementing OCT's structural signal with molecular imaging requires strategies to simultaneously detect multiple exogenous contrast agents with high specificity in tissue. Specific detection of molecular probes enables the parallel visualization of physiological, cellular, and molecular processes. Here we demonstrate that, by combining PS-OCT and spectral contrast (SC)-OCT measurements, we can distinguish signatures of different gold nanobipyramids (GNBPs) in lymphatic vessels from the surrounding tissue and blood vessels in live mouse models. This technique could well be extended to other anisotropic nanoparticle-based OCT contrast agents and presents significant progress toward enabling OCT molecular imaging.

Transient-mode photothermal optical coherence tomography.

Photothermal optical coherence tomography (PT-OCT) is an emerging extension of OCT, which forms images based on both scattering and absorption of light. The speed of PT-OCT, however, has been limited by the necessity for lock-in detection with extensive temporal sampling of the sample's PT response. Here, we demonstrate transient-mode PT-OCT (TM-PT-OCT), which increases the effective A-line rate by orders of magnitude from 10-100 Hz to 1.5-7.5 kHz, by interrogating the sample's transient thermal response to a single diode laser pulse. Functional imaging of moving samples with TM-PT-OCT at video rate is also presented. This significant improvement in imaging speed is expected to open the door for downstream integration of PT-OCT in clinical systems for in vivo imaging.

Polarimetric Signatures of Coronary Thrombus in Patients With Acute Coronary Syndrome.

BACKGROUND: Intravascular polarization-sensitive optical frequency domain imaging (PS-OFDI) offers a novel approach to measure tissue birefringence, which is elevated in collagen and smooth muscle cells, that in turn plays a critical role in healing coronary thrombus (HCT). This study aimed to quantitatively assess polarization properties of coronary fresh and organizing thrombus with PS-OFDI in patients with acute coronary syndrome (ACS).Methods and Results:The POLARIS-I prospective registry enrolled 32 patients with ACS. Pre-procedural PS-OFDI pullbacks using conventional imaging catheters revealed 26 thrombus-regions in 21 patients. Thrombus was manually delineated in conventional OFDI cross-sections separated by 0.5 mm and categorized into fresh thrombus caused by plaque rupture, stent thrombosis, or erosion in 18 thrombus-regions (182 frames) or into HCT for 8 thrombus-regions (141 frames). Birefringence of coronary thrombus was compared between the 2 categories. Birefringence in HCTs was significantly higher than in fresh thrombus (∆n=0.47 (0.37-0.72) vs. ∆n=0.25 (0.17-0.29), P=0.007). In a subgroup analysis, when only using thrombus-regions from culprit lesions, ischemic time was a significant predictor for birefringence (ß (∆n)=0.001 per hour, 95% CI [0.0002-0.002], P=0.023). CONCLUSIONS: Intravascular PS-OFDI offers the opportunity to quantitatively assess the polarimetric properties of fresh and organizing coronary thrombus, providing new insights into vascular healing and plaque stability.

Single-shot depth profiling by spatio-temporal encoding with a multimode fiber.

Computational imaging with random encoding patterns obtained by scattering of light in complex media has enabled simple imaging systems with compelling performance. Here, we extend this concept to axial reflectivity profiling using spatio-temporal coupling of broadband light in a multimode fiber (MMF) to generate the encoding functions. Interference of light transmitted through the MMF with a sample beam results in path-length-specific patterns that enable computational reconstruction of the axial sample reflectivity profile from a single camera snapshot. Leveraging the versatile nature of MMFs, we demonstrate depth profiling with bandwidth-limited axial resolution of 13.4 µm over a scalable sensing range reaching well beyond one centimeter.

In vivo imaging of the depth-resolved optic axis of birefringence in human skin.

Recent progress has enabled the reconstruction of the local (i.e., depth-resolved) optic axis (OAx) of biological tissue from measurements made with polarization-sensitive optical coherence tomography (PS-OCT). Here we demonstrate local OAx imaging in healthy human skin in vivo. The images reveal dense, weaving patterns that are imperceptible in OCT intensity tomograms or conventional PS-OCT metrics and that suggest a mesh-like tissue organization, consistent with the morphology of dermal collagen. Using co-registered polarization-sensitive optical coherence microscopy, we furthermore investigated the impact of spatial resolution on the recovered OAx patterns and confirmed their consistency. OAx orientation as a contrast mechanism merits further exploration for applications in dermatology.

Linear-in-wavenumber actively-mode-locked wavelength-swept laser.

We report on an akinetic actively-mode-locked wavelength-swept laser (ASL) with a sweep that is highly linear in wavenumber. By tailoring the drive waveform of the intracavity modulator, the wavenumber sweep was further linearized to enable high fidelity frequency-domain interferometric ranging without resampling of the acquired data. Used for catheter-based optical coherence tomography, the ASL showed comparable imaging performance to a state-of-the-art polygon-based wavelength-swept source at a matching sweep rate of 103.6 kHz, a duty cycle of 95%, and a bandwidth of 100 nm, centered at 1330 nm.

Robust wavenumber and dispersion calibration for Fourier-domain optical coherence tomography.

Many Fourier-domain optical coherence tomography (FD-OCT) systems sample the interference fringes with a non-uniform wavenumber (k) interval, introducing a chirp to the signal that depends on the path length difference underlying each fringe. A dispersion imbalance between sample and reference arms also generates a chirp in the fringe signal which, in contrast, is independent of depth. Fringe interpolation to obtain a signal linear in k and compensate dispersion imbalance is critical to achieving bandwidth-limited axial resolution. In this work, we propose an optimization-based algorithm to perform robust and automated calibration of FD-OCT systems, recovering both the interpolation function and the dispersion imbalance. Our technique relies on the fact that the unique function that correctly linearizes the fringe data in k space produces a depth-independent chirp. The calibration procedure requires experimental data corresponding to a single reflector at various depth locations, which can easily be obtained by acquiring data while moving a sample mirror in depth. We have tested both spectral domain OCT and swept source OCT systems with various nonlinearities. Results indicate that the proposed calibration method has excellent performance on a wide range of data sets and enables nearly constant resolution at all imaging depths. An implementation of the algorithm is available online.

Extended bandwidth wavelength swept laser source for high resolution optical frequency domain imaging.

Improving the axial resolution by providing wider bandwidth wavelength swept lasers remains an important issue for optical frequency domain imaging (OFDI). Here, we demonstrate a wide tuning range, all-fiber wavelength swept laser at a center wavelength of 1250 nm by combining two ring cavities that share a single Fabry-Perot tunable filter. The two cavities contain semiconductor optical amplifiers with central wavelengths of 1190 nm and 1292 nm, respectively. To avoid disturbing interference effects in the overlapping spectral region, we modulated the amplifiers in order to obtain consecutive wavelength sweeps in the two spectral regions. The two sweeps were fused together in post-processing to achieve a total scanning range of 223 nm, corresponding to 3.3 µm axial resolution in air. We confirm improved image quality and reduced speckle size in tomograms of swine esophagus ex vivo, and human skin and nailbed in vivo.

Longitudinal three-dimensional visualisation of autoimmune diabetes by functional optical coherence imaging.

AIMS/HYPOTHESIS: It is generally accepted that structural and functional quantitative imaging of individual islets would be beneficial to elucidate the pathogenesis of type 1 diabetes. We here introduce functional optical coherence imaging (FOCI) for fast, label-free monitoring of beta cell destruction and associated alterations of islet vascularisation. METHODS: NOD mouse and human islets transplanted into the anterior chamber of the eye (ACE) were imaged with FOCI, in which the optical contrast of FOCI is based on intrinsic variations of the index of refraction resulting in a faster tomographic acquisition. In addition, the phase sensitivity allows simultaneous label-free acquisition of vascularisation. RESULTS: We demonstrate that FOCI allows longitudinal quantification of progressive autoimmune insulitis, including the three-dimensional quantification of beta cell volume, inflammation and vascularisation. The substantially increased backscattering of islets is dominated by the insulin-zinc nanocrystals in the beta cell granules. This translates into a high specificity for the functional beta cell volume of islets. Applying FOCI to a spontaneous mouse model of type 1 diabetes, we quantify the modifications of the pancreatic microvasculature accompanying the progression of diabetes and reveal a strong correlation between increasing insulitis and density of the vascular network of the islet. CONCLUSIONS/INTERPRETATION: FOCI provides a novel imaging technique for investigating functional and structural diabetes-induced alterations of the islets. The label-free detection of beta cell volume and infiltration together with vascularisation offers a unique extension to study ACE-transplanted human islets. These results are contributing to a deeper understanding of human islet transplant rejection and label-free in vivo monitoring of drug efficacy.

Ultrahigh-resolution optical coherence elastography.

Visualizing stiffness within the local tissue environment at the cellular and subcellular level promises to provide insight into the genesis and progression of disease. In this Letter, we propose ultrahigh-resolution optical coherence elastography (UHROCE), and demonstrate 3D imaging of local axial strain of tissues undergoing compressive loading. We combine optical coherence microscopy (OCM) and phase-sensitive detection of local tissue displacement to produce strain elastograms with resolution (x,y,z) of 2×2×15 µm. We demonstrate this performance on a freshly excised mouse aorta and reveal the mechanical heterogeneity of vascular smooth muscle cells and elastin sheets, otherwise unresolved in a typical, lower resolution optical coherence elastography (OCE) system.

Single input state, single-mode fiber-based polarization-sensitive optical frequency domain imaging by eigenpolarization referencing.

Fiber-based polarization-sensitive optical frequency domain imaging is more challenging than free-space implementations. Using multiple input states, fiber-based systems provide sample birefringence information with the benefit of a flexible sample arm but come at the cost of increased system and acquisition complexity, and either reduce acquisition speed or require increased acquisition bandwidth. Here we show that with the calibration of a single polarization state, fiber-based configurations can approach the conceptual simplicity of traditional free-space configurations. We remotely control the polarization state of the light incident at the sample using the eigenpolarization states of a wave plate as a reference, and determine the Jones matrix of the output fiber. We demonstrate this method for polarization-sensitive imaging of biological samples.

Degree of polarization (uniformity) and depolarization index: unambiguous depolarization contrast for optical coherence tomography.

The degree of polarization (uniformity) has attracted increased interest as a functional contrast in optical coherence tomography (OCT). However, its computation from a single polarization state suggests an ambiguity that is strongly dependent on a sample's orientation. We here propose an improved metric to present depolarization with respect to the optical system rather than the propagating field. Using numerical simulations and optical frequency domain imaging, we evaluate the conventional DOP(U) for different polarization states and compare its performance with the unambiguous depolarization index.

An automated image processing method to quantify collagen fibre organization within cutaneous scar tissue.

Standard approaches to evaluate scar formation within histological sections rely on qualitative evaluations and scoring, which limits our understanding of the remodelling process. We have recently developed an image analysis technique for the rapid quantification of fibre alignment at each pixel location. The goal of this study was to evaluate its application for quantitatively mapping scar formation in histological sections of cutaneous burns. To this end, we utilized directional statistics to define maps of fibre density and directional variance from Masson's trichrome-stained sections for quantifying changes in collagen organization during scar remodelling. Significant increases in collagen fibre density are detectable soon after burn injury in a rat model. Decreased fibre directional variance in the scar was also detectable between 3 weeks and 6 months after injury, indicating increasing fibre alignment. This automated analysis of fibre organization can provide objective surrogate endpoints for evaluating cutaneous wound repair and regeneration.

Quantitative technique for robust and noise-tolerant speed measurements based on speckle decorrelation in optical coherence tomography.

Intensity-based techniques in optical coherence tomography (OCT), such as those based on speckle decorrelation, have attracted great interest for biomedical and industrial applications requiring speed or flow information. In this work we present a rigorous analysis of the effects of noise on speckle decorrelation, demonstrate that these effects frustrate accurate speed quantitation, and propose new techniques that achieve quantitative and repeatable measurements. First, we derive the effect of background noise on the speckle autocorrelation function, finding two detrimental effects of noise. We propose a new autocorrelation function that is immune to the main effect of background noise and permits quantitative measurements at high and moderate signal-to-noise ratios. At the same time, this autocorrelation function is able to provide motion contrast information that accurately identifies areas with movement, similar to speckle variance techniques. In order to extend the SNR range, we quantify and model the second effect of background noise on the autocorrelation function through a calibration. By obtaining an explicit expression for the decorrelation time as a function of speed and diffusion, we show how to use our autocorrelation function and noise calibration to measure a flowing liquid. We obtain accurate results, which are validated by Doppler OCT, and demonstrate a very high dynamic range (> 600 mm/s) compared to that of Doppler OCT (±25 mm/s). We also derive the behavior for low flows, and show that there is an inherent non-linearity in speed measurements in the presence of diffusion due to statistical fluctuations of speckle. Our technique allows quantitative and robust measurements of speeds using OCT, and this work delimits precisely the conditions in which it is accurate.

All-fiber wavelength swept ring laser based on Fabry-Perot filter for optical frequency domain imaging.

Innovations in laser engineering have yielded several novel configurations for high repetition rate, broad sweep range, and long coherence length wavelength swept lasers. Although these lasers have enabled high performance frequency-domain optical coherence tomography, they are typically complicated and costly and many require access to proprietary materials or devices. Here, we demonstrate a simplified ring resonator configuration that is straightforward to construct from readily available materials at a low total cost. It was enabled by an insight regarding the significance of isolation against bidirectional operation and by configuring the sweep range of the intracavity filter to exceed its free spectral range. The design can easily be optimized to meet a range of operating specifications while yielding robust and stable performance. As an example, we demonstrate 240 kHz operation with 125 nm sweep range and >70 mW of average output power and demonstrate high quality frequency domain OCT imaging. The complete component list and directions for assembly of the laser are posted on-line at www.octresearch.org.

Practical decomposition for physically admissible differential Mueller matrices.

The differential Mueller matrix expresses the local action of an optical medium on the evolution of a propagating electromagnetic field, including partially coherent and partially polarized waves. Here, we present a derivation of the differential Mueller matrix from the canonical form of Type I Mueller matrices without making use of the exponential generators of uniform media. We demonstrate how to practically obtain this parameterization numerically using an eigenvalue decomposition and find validity criteria to ensure that the matrix satisfies the constraints of a physical system. This provides a convenient tool-set to investigate depolarization effects and extends previous treatments of the differential Mueller matrix formalism.

Artificial neural network for enhancing signal-to-noise ratio and contrast in photothermal optical coherence tomography.

Optical coherence tomography (OCT) is a medical imaging method that generates micron-resolution 3D volumetric images of tissues in-vivo. Photothermal (PT)-OCT is a functional extension of OCT with the potential to provide depth-resolved molecular information complementary to the OCT structural images. PT-OCT typically requires long acquisition times to measure small fluctuations in the OCT phase signal. Here, we use machine learning with a neural network to infer the amplitude of the photothermal phase modulation from a short signal trace, trained in a supervised fashion with the ground truth signal obtained by conventional reconstruction of the PT-OCT signal from a longer acquisition trace. Results from phantom and tissue studies show that the developed network improves signal to noise ratio (SNR) and contrast, enabling PT-OCT imaging with short acquisition times and without any hardware modification to the PT-OCT system. The developed network removes one of the key barriers in translation of PT-OCT (i.e., long acquisition time) to the clinic.