Aging of deep venous thrombosis in-vivo using polarization sensitive optical coherence tomography.

Deep venous thrombosis (DVT) is a medical condition with significant post-event morbidity and mortality coupled with limited treatment options. Treatment strategy and efficacy are highly dependent on the structural composition of the thrombus, which evolves over time from initial formation and is currently unevaluable with standard clinical testing. Here, we investigate the use of intravascular polarization-sensitive optical coherence tomography (PS-OCT) to assess thrombus morphology and composition in a rat DVT model in-vivo, including changes that occur over the thrombus aging process. PS-OCT measures tissue birefringence, which provides contrast for collagen and smooth muscle cells that are present in older, chronic clots. Thrombi in the inferior vena cava of two cohorts of rats were imaged in-vivo with intravascular PS-OCT at 24 hours (acute, nrats = 3, 73 cross-sections) or 28 days (chronic, nrats = 4, 41 cross-sections) after thrombus formation. Co-registered histology was labelled by an independent pathologist to establish ground-truth clot composition. Automated analysis of OCT cross-sectional images differentiated acute and chronic thrombi with 97.6% sensitivity and 98.6% specificity using a linear discriminant model comprised of both polarization and conventional OCT metrics. These results support PS-OCT as a highly sensitive imaging modality for the assessment of DVT composition to differentiate acute and chronic thrombi. Intravascular PS-OCT imaging could be integrated with advanced catheter-based treatment strategies and serve to guide therapeutic decision-making and deployment, by offering an accurate assessment of DVT patients in real time.

Segmentation of anatomical layers and imaging artifacts in intravascular polarization sensitive optical coherence tomography using attending physician and boundary cardinality losses.

Intravascular ultrasound and optical coherence tomography are widely available for assessing coronary stenoses and provide critical information to optimize percutaneous coronary intervention. Intravascular polarization-sensitive optical coherence tomography (PS-OCT) measures the polarization state of the light scattered by the vessel wall in addition to conventional cross-sectional images of subsurface microstructure. This affords reconstruction of tissue polarization properties and reveals improved contrast between the layers of the vessel wall along with insight into collagen and smooth muscle content. Here, we propose a convolutional neural network model, optimized using two new loss terms (Boundary Cardinality and Attending Physician), that takes advantage of the additional polarization contrast and classifies the lumen, intima, and media layers in addition to guidewire and plaque shadows. Our model segments the media boundaries through fibrotic plaques and continues to estimate the outer media boundary behind shadows of lipid-rich plaques. We demonstrate that our multi-class classification model outperforms existing methods that exclusively use conventional OCT data, predominantly segment the lumen, and consider subsurface layers at most in regions of minimal disease. Segmentation of all anatomical layers throughout diseased vessels may facilitate stent sizing and will enable automated characterization of plaque polarization properties for investigation of the natural history and significance of coronary atheromas.

Probabilistic volumetric speckle suppression in OCT using deep learning.

We present a deep learning framework for volumetric speckle reduction in optical coherence tomography (OCT) based on a conditional generative adversarial network (cGAN) that leverages the volumetric nature of OCT data. In order to utilize the volumetric nature of OCT data, our network takes partial OCT volumes as input, resulting in artifact-free despeckled volumes that exhibit excellent speckle reduction and resolution preservation in all three dimensions. Furthermore, we address the ongoing challenge of generating ground truth data for supervised speckle suppression deep learning frameworks by using volumetric non-local means despeckling-TNode to generate training data. We show that, while TNode processing is computationally demanding, it serves as a convenient, accessible gold-standard source for training data; our cGAN replicates efficient suppression of speckle while preserving tissue structures with dimensions approaching the system resolution of non-local means despeckling while being two orders of magnitude faster than TNode. We demonstrate fast, effective, and high-quality despeckling of the proposed network in different tissue types acquired with three different OCT systems compared to existing deep learning methods. The open-source nature of our work facilitates re-training and deployment in any OCT system with an all-software implementation, working around the challenge of generating high-quality, speckle-free training data.

Single-input polarization-sensitive optical coherence tomography through a catheter.

Intravascular polarimetry with catheter-based polarization-sensitive optical coherence tomography (PS-OCT) complements the high-resolution structural tomograms of OCT with morphological contrast available through polarimetry. Its clinical translation has been complicated by the need for modification of conventional OCT hardware to enable polarimetric measurements. Here, we present a signal processing method to reconstruct the polarization properties of tissue from measurements with a single input polarization state, bypassing the need for modulation or multiplexing of input states. Our method relies on a polarization symmetry intrinsic to round-trip measurements and uses the residual spectral variation of the polarization states incident on the tissue to avoid measurement ambiguities. We demonstrate depth-resolved birefringence and optic axis orientation maps reconstructed from in-vivo data of human coronary arteries. We validate our method through comparison with conventional dual-input state measurements and find a mean cumulative retardance error of 13.2deg without observable bias. The 95% limit of agreement between depth-resolved birefringence is 2.80 · 10-4, which is less than the agreement between two repeat pullbacks of conventional PS-OCT (3.14 · 10-4), indicating that the two methods can be used interchangeably. The hardware simplification arising from using a single input state may be decisive in realizing the potential of polarimetric measurements for assessing coronary atherosclerosis in clinical practice.

Mapping optical scattering properties to physical particle information in singly and multiply scattering samples.

Optical coherence tomography (OCT) leverages light scattering by biological tissues as endogenous contrast to form structural images. Light scattering behavior is dictated by the optical properties of the tissue, which depend on microstructural details at the cellular or sub-cellular level. Methods to measure these properties from OCT intensity data have been explored in the context of a number of biomedical applications seeking to access this sub-resolution tissue microstructure and thereby increase the diagnostic impact of OCT. Most commonly, the optical attenuation coefficient, an analogue of the scattering coefficient, has been used as a surrogate metric linking OCT intensity to subcellular particle characteristics. To record attenuation coefficient data that is accurately representative of the underlying physical properties of a given sample, it is necessary to account for the impact of the OCT imaging system itself on the distribution of light intensity in the sample, including the numerical aperture (NA) of the system and the location of the focal plane with respect to the sample surface, as well as the potential contribution of multiple scattering to the reconstructed intensity signal. Although these considerations complicate attenuation coefficient measurement and interpretation, a suitably calibrated system may potentiate a powerful strategy for gaining additional information about the scattering behavior and microstructure of samples. In this work, we experimentally show that altering the OCT system geometry minimally impacts measured attenuation coefficients in samples presumed to be singly scattering, but changes these measurements in more highly scattering samples. Using both depth-resolved attenuation coefficient data and layer-resolved backscattering coefficients, we demonstrate the retrieval of scattering particle diameter and concentration in tissue-mimicking phantoms, and the impact of presumed multiple scattering on these calculations. We further extend our approach to characterize a murine brain tissue sample and highlight a tumor-bearing region based on increased scattering particle density. Through these methods, we not only enhance conventional OCT attenuation coefficient analysis by decoupling the independent effects of particle size and concentration, but also discriminate areas of strong multiple scattering through minor changes to system topology to provide a framework for assessing the accuracy of these measurements.

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.

Efficient dispersion modeling in optical multimode fiber.

Dispersion remains an enduring challenge for the characterization of wavelength-dependent transmission through optical multimode fiber (MMF). Beyond a small spectral correlation width, a change in wavelength elicits a seemingly independent distribution of the transmitted field. Here we report on a parametric dispersion model that describes mode mixing in MMF as an exponential map and extends the concept of principal modes to describe the fiber's spectrally resolved transmission matrix (TM). We present computational methods to fit the model to measurements at only a few, judiciously selected, discrete wavelengths. We validate the model in various MMF and demonstrate an accurate estimation of the full TM across a broad spectral bandwidth, approaching the bandwidth of the best-performing principal modes, and exceeding the original spectral correlation width by more than two orders of magnitude. The model allows us to conveniently study the spectral behavior of principal modes, and obviates the need for dense spectral measurements, enabling highly efficient reconstruction of the multispectral TM of MMF.

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.

Measuring collagen injury depth for burn severity determination using polarization sensitive optical coherence tomography.

Determining the optimal treatment course for a dermatologic burn wound requires knowledge of the wound's severity, as quantified by the depth of thermal damage. In current clinical practice, burn depth is inferred based exclusively on superficial visual assessment, a method which is subject to substantial error rates in the classification of partial thickness (second degree) burns. Here, we present methods for direct, quantitative determination of the depth extent of injury to the dermal collagen matrix using polarization-sensitive optical coherence tomography (PS-OCT). By visualizing the depth-dependence of the degree of polarization of light in the tissue, rather than cumulative retardation, we enable direct and volumetric assessment of local collagen status. We further augment our PS-OCT measurements by visualizing adnexal structures such as hair follicles to relay overall dermal viability in the wounded region. Our methods, which we have validated ex vivo with matched histology, offer an information-rich tool for precise interrogation of burn wound severity and healing potential in both research and clinical settings.

Confocal 3D reflectance imaging through multimode fiber without wavefront shaping.

Imaging through optical multimode fiber (MMF) has the potential to enable hair-thin endoscopes that reduce the invasiveness of imaging deep inside tissues and organs. Active wavefront shaping and fluorescent labeling have recently been exploited to overcome modal scrambling and enable MMF imaging. Here, we present a computational approach that circumvents the need for active wavefront control and exogenous fluorophores. We demonstrate the reconstruction of depth-gated confocal images through MMF using a raster-scanned, focused input illumination at the fiber proximal end. We show the compatibility of this approach with quantitative phase, dark-field, and polarimetric imaging. Computational imaging through MMF opens a new pathway for minimally invasive imaging in medical diagnosis and biological investigations.

Nanosecond SRS fiber amplifier for label-free near-infrared photoacoustic microscopy of lipids.

Near-infrared photoacoustics receives increasing interest as an intravital modality to sense key biomolecules. One of the most central types of biomolecules of interest are lipids as they constitute essential bio-hallmarks of cardiovascular and metabolic diseases and their in-vivo detection holds insightful information about disease progression and treatment monitoring. However, the full potential of near-infrared photoacoustic for high-resolution and high-sensitivity biomedical studies of lipids has so far not been exploited due a lack of appropriate excitation sources delivering short-pulses at high-repetition-rate, high-pulse-energy, and wavelength around 1200 nm. Here, we demonstrate a custom-built SRS fiber amplifier that provides optical excitations at 1192.8 nm, repetition rates of 200 kHz, pulse durations below 2 ns, and pulse energies beyond 5 µJ. We capitalize on the performance of our excitation source and show near-infrared photoacoustics resolving intrinsic lipid contrast in biomedically relevant specimens ranging from single cells to lipid-rich tissue with subcellular resolution.

Layer-based, depth-resolved computation of attenuation coefficients and backscattering fractions in tissue using optical coherence tomography.

Structural optical coherence tomography (OCT) images of tissue stand to benefit from greater functionalization and quantitative interpretation. The OCT attenuation coefficient µ, an analogue of the imaged sample's scattering coefficient, offers potential functional contrast based on the relationship of µ to sub-resolution physical properties of the sample. Attenuation coefficients are computed either by fitting a representative µ over several depth-wise pixels of a sample's intensity decay, or by using previously-developed depth-resolved attenuation algorithms by Girard et al. [Invest. Ophthalmol. Vis. Sci.52, 7738 (2011). 10.1167/iovs.10-6925] and Vermeer et al. [Biomed. Opt. Express5, 322 (2014). 10.1364/BOE.5.000322]. However, the former method sacrifices axial information in the tomogram, while the latter relies on the stringent assumption that the sample's backscattering fraction, another optical property, does not vary along depth. This assumption may be violated by layered tissues commonly observed in clinical imaging applications. Our approach preserves the full depth resolution of the attenuation map but removes its dependence on backscattering fraction by performing signal analysis inside individual discrete layers over which the scattering properties (e.g., attenuation and backscattering fraction) vary minimally. Although this approach necessitates the detection of these layers, it removes the constant-backscattering-fraction assumption that has constrained quantitative attenuation coefficient analysis in the past, and additionally yields a layer-resolved backscattering fraction, providing complementary scattering information to the attenuation coefficient. We validate our approach using automated layer detection in layered phantoms, for which the measured optical properties were in good agreement with theoretical values calculated with Mie theory, and show preliminary results in tissue alongside corresponding histological analysis. Together, accurate backscattering fraction and attenuation coefficient measurements enable the estimation of both particle density and size, which is not possible from attenuation measurements alone. We hope that this improvement to depth-resolved attenuation coefficient measurement, augmented by a layer-resolved backscattering fraction, will increase the diagnostic power of quantitative OCT imaging.

Whole Murine Brain Imaging Based on Optical Elastic Scattering.

Imaging whole brains is one of the central efforts of biophotonics. While the established imaging modalities used in radiology, such as MRI and CT, have enabled in vivo investigations of various cognitive and affective processes, the prevailing resolution of one-cubic-millimeter has limited their use in studying the "ground-truth" of neuronal activities. On the other hand, electron microscopy (EM) visualizes the finest anatomic structures at a resolution of around 30 nm. However, the extensive tissue preparation process and the required large-scale morphological reconstruction restrict this method to small sample volumes. Light microscopy (LM) has the potential to bridge the above two spatial scales, with a resolution ranging from a few hundred nanometers to a few micrometers. Recent advances in tissue clearing have paved the way for optical investigation of large intact tissue volumes. However, most of these LM studies rely on fluorescence-a nonlinear optical process to provide contrast. This chapter introduces an alternative type of LM that is solely based on a linear optical process-elastic scattering, which has some unique advantages over conventional LM methods for the investigation of large-scale biological systems, such as intact murine brains. Here, we will first lay out the background and the motivation of developing this scattering-based method. Then, the basic principle of this approach will be introduced, including controlling tissue scattering and coherent imaging. Next, we explore current implementation and practical considerations. Up-to-date results and the utility of this method will also be demonstrated. Finally, we discuss current limitations and future directions in this promising field.

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.

Retinal imaging in Alzheimer's and neurodegenerative diseases.

In the last 20 years, research focused on developing retinal imaging as a source of potential biomarkers for Alzheimer's disease and other neurodegenerative diseases, has increased significantly. The Alzheimer's Association and the Alzheimer's & Dementia: Diagnosis, Assessment, Disease Monitoring editorial team (companion journal to Alzheimer's & Dementia) convened an interdisciplinary discussion in 2019 to identify a path to expedite the development of retinal biomarkers capable of identifying biological changes associated with AD, and for tracking progression of disease severity over time. As different retinal imaging modalities provide different types of structural and/or functional information, the discussion reflected on these modalities and their respective strengths and weaknesses. Discussion further focused on the importance of defining the context of use to help guide the development of retinal biomarkers. Moving from research to context of use, and ultimately to clinical evaluation, this article outlines ongoing retinal imaging research today in Alzheimer's and other brain diseases, including a discussion of future directions for this area of study.

Computational adaptive optics in phase-unstable optical coherence tomography.

We present a scheme for correction of x-y-separable aberrations in optical coherence tomography (OCT) designed to work with phase unstable systems with no hardware modifications. Our approach, termed SHARP, is based on computational adaptive optics and numerical phase correction and follows from the fact that local phase stability is sufficient for the deconvolution of optical aberrations. We demonstrate its applicability in a raster-scan polygon-laser OCT system with strong phase-jitter noise, achieving successful refocusing at depths up to 4 times the Rayleigh range. We also present in vivo endoscopic and ex vivo anterior segment OCT data, showing significant enhancement of image quality, particularly when combining SHARP results with a resolution-preserving despeckling technique like TNode.

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.

Intravascular Polarimetry: Clinical Translation and Future Applications of Catheter-Based Polarization Sensitive Optical Frequency Domain Imaging.

Optical coherence tomography (OCT) and optical frequency domain imaging (OFDI) visualize the coronary artery wall and plaque morphology in great detail. The advent of these high-resolution intracoronary imaging modalities has propelled our understanding of coronary atherosclerosis and provided enhanced guidance for percutaneous coronary intervention. Yet, the lack of contrast between distinct tissue types and plaque compositions impedes further elucidation of the complex mechanisms that contribute to acute coronary syndrome (ACS) and hinders the prospective identification of plaques susceptible to rupture. Intravascular polarimetry with polarization-sensitive OFDI measures polarization properties of the coronary arterial wall using conventional intravascular imaging catheters. The quantitative polarization metrics display notable image contrast between several relevant coronary plaque microstructures that are difficult to identify with conventional OCT and OFDI. Tissues rich in collagen and smooth muscle cells exhibit birefringence, while lipid and macrophages cause depolarization. In this review, we describe the basic principles of intravascular polarimetry, discuss the interpretation of the polarization signatures, and outline promising avenues for future research and clinical implications.