A turnkey phase-sensitive swept-source optical coherence tomography (OCT) system requires stabilization of both the swept source and main signal interferometer. We have provided stable clocking and triggering of a 1050 nm swept micro-electromechanical system (MEMS) vertical cavity surface emitting laser (VCSEL) by mounting a solid clock etalon and volume Bragg grating optical trigger inside a temperature-stabilized butterfly package. A photonic integrated circuit (PIC) provides a phase-stable main interferometer near the sample. This system is only sensitive to the motion of the sample, not to the fibers leading to and from the PIC. We provide optical frequency domain reflectivity measurements of the PIC performance. We also demonstrate OCT imaging as well as static and dynamic phase measurements of a Kindle e-book reader electrophoretic display.

The flexible membranes used in MEMS tunable VCSELs are so small and light that thermally induced vibrations can impact laser performance. We measure the thermal vibration spectrum of such a membrane showing peaks at the spatial vibration mode resonant frequencies of the membrane/plate. These vibrations result in a theoretical floor to the linewidth of the VCSEL. Frequency domain LiDAR and optical coherence tomography systems can get around this thermal linewidth limit with adequate clock measurement and processing. Essentially an OCT/LiDAR sweep with a concomitantly measured clock is a feed-forward linewidth reduction scheme. This can be achieved because the membrane resonances are relatively low frequency. LiDAR ranging out to 9 meters has been demonstrated with a resolution of 13 μm, close to the transform limit for the 70 nm sampling range.

Low NEP balanced receivers generally do not result in better sensitivity in a shot-noise-limited swept source OCT system. However, there is an advantage if RIN is significant. A lower NEP, even in the shot-noise-limited case, does allow for lower reference arm powers. This, in turn, reduces fixed pattern artifact signals caused by stray optical component reflections inside a swept source laser cavity. An NEP reduction of √10 allows the reference power to be reduced 10 dB while maintaining SNR. This reduces the pattern to noise ratio by 10 dB because pattern noise is directly detected (20 dB scaling per decade reference power), whereas the image signal is heterodyne detected (10 dB scaling per decade of reference power). We present sensitivity and fixed pattern measurements taken with six commercial balanced receivers, including an APD receiver. We also present an NEP survey of 23 commercial receivers over a wide range of bandwidths and transimpedances.

Side lobe artifacts on point spread functions can be traced back to (1) fringe visibility variation across the spectrum, (2) errors in sampling instances, and (3) window functions. We demonstrate signal processing methods for correcting for all three of these issues. These methods require a system calibration step. If the systems slowly age, the recalibration step could be performed in the field with a fixtured target.

A 1060 nm optically pumped tunable VCSEL was formed from an InGaAs/AlGaAs/GaAs half-VCSEL bonded to a MEMS movable mirror on a silicon substrate. The VCSEL was co-packaged in a 14-pin butterfly module with an 825 nm pump laser and a 1060 nm semiconductor optical amplifier. The co-packaged device exhibited shot-noise-limited sensitivity with up to 50 mW output power and 75 nm tunability. Ophthalmic OCT, especially whole-eye imaging and ocular biometry, is considered the primary application of this device. However, we have also investigated LiDAR to greater than 10 meter ranges with non-mechanical beam steering through angular diffraction from a grating. A new generation of photonic integrated circuit LiDARs work this way and we have investigated the depth resolution limitations due to time dispersion from the grating. Distributed fiber temperature sensing was also demonstrated.

Cystic fibrosis (CF) is a genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. Although impairment of mucociliary clearance contributes to severe morbidity and mortality in people with CF, a clear understanding of the pathophysiology is lacking. This is, in part, due to the absence of clinical imaging techniques capable of capturing CFTR-dependent functional metrics at the cellular level. Here, we report the clinical translation of a 1-μm resolution micro-optical coherence tomography (μOCT) technology to quantitatively characterize the functional microanatomy of human upper airways. Using a minimally invasive intranasal imaging approach, we performed a clinical study on age- and sex-matched CF and control groups. We observed delayed mucociliary transport rate at the cellular level, depletion of periciliary liquid layer, and prevalent loss of ciliation in subjects with CF. Distinctive morphological differences in mucus and various forms of epithelial injury were also revealed by μOCT imaging and had prominent effects on the mucociliary transport apparatus. Elevated mucus reflectance intensity in CF, a proxy for viscosity in situ, had a dominant effect. These results demonstrate the utility of μOCT to determine epithelial function and monitor disease status of CF airways on a per-patient basis, with applicability for other diseases of mucus clearance.

Near infrared imaging methods, including optical coherence tomography (OCT), have shown promise in assessing activity of enamel caries lesions. We present initial results in ex-vivo dental samples with a polarization diversity-detection OCT system complete with an integrated dental handpiece. Built around an Axsun Technologies swept source engine, images B-scans 12(x) by 7(z) mm at over 100 fps with 66um transverse and 7um depth resolution. Internal normal dental structure and lesions were imaged and compared with micro-CT. We demonstrate that the Axsun OCT prototype can detect the penetration of occlusal lesions to dentin and measure the penetration of approximal lesions when imaging from the occlusal surface.

Neutrophil breach of the mucosal surface is a common pathological consequence of infection. We present an advanced co-culture model to explore neutrophil transepithelial migration utilizing airway mucosal barriers differentiated from primary human airway basal cells and examined by advanced imaging. Human airway basal cells were differentiated and cultured at air-liquid interface (ALI) on the underside of 3 µm pore-sized transwells, compatible with the study of transmigrating neutrophils. Inverted ALIs exhibit beating cilia and mucus production, consistent with conventional ALIs, as visualized by micro-optical coherence tomography (µOCT). µOCT is a recently developed imaging modality with the capacity for real time two- and three-dimensional analysis of cellular events in marked detail, including neutrophil transmigratory dynamics. Further, the newly devised and imaged primary co-culture model recapitulates key molecular mechanisms that underlie bacteria-induced neutrophil transepithelial migration previously characterized using cell line-based models. Neutrophils respond to imposed chemotactic gradients, and migrate in response to Pseudomonas aeruginosa infection of primary ALI barriers through a hepoxilin A3-directed mechanism. This primary cell-based co-culture system combined with µOCT imaging offers significant opportunity to probe, in great detail, micro-anatomical and mechanistic features of bacteria-induced neutrophil transepithelial migration and other important immunological and physiological processes at the mucosal surface.

The human respiratory system is protected by a defense mechanism termed mucociliary clearance (MCC). Deficiency in MCC leads to respiratory obstruction and pulmonary infection, which often are the main causes of morbidity and mortality in diseases such as cystic fibrosis and chronic obstructive pulmonary disease (COPD). Studying key parameters that govern MCC, including ciliary beat frequency, velocity and volume of airway mucus transport, as well as periciliary liquid layer thickness are therefore of great importance in understanding human respiratory health. However, direct, in vivo visualization of ciliary function and MCC has been challenging, hindering the diagnosis of disease pathogenesis and mechanistic evaluation of novel therapeutics. Our laboratory has previously developed a 1-µm resolution optical coherence tomography method, termed Micro-OCT, which is a unique tool for visualizing the spatiotemporal features of ciliary function and MCC. We have previously described the design of a flexible 2.5 mm Micro-OCT probe that is compatible with standard flexible bronchoscopes. This device utilizes a common-path interferometer and annular sample arm apodization to attain a sharply focused spot over an extended depth of focus. Here, we present the most recent iteration of this probe and demonstrate its imaging performance in a mouse trachea tissue culture model. In addition, we have developed an ergonomic assembly for attaching the probe to a standard bronchoscope. The ergonomic assembly fixes the Micro-OCT probe’s within the bronchoscope and contains a means transducing linear motion through the sheath so that the Micro-OCT beam can be scanned along the trachea. We have tested the performance of these devices for Micro-OCT imaging in an anatomically correct model of the human airway. Future studies are planned to use this technology to conduct Micro-OCT in human trachea and bronchi in vivo.

We report the design and fabrication of a flexible, longitudinally scanning high-resolution micro-optical coherence tomography (μOCT) endobronchial probe, optimized for micro-anatomical imaging in airways. The 2.4 mm diameter and flexibility of the probe allows it to be inserted into the instrument channel of a standard bronchoscope, enabling real-time video guidance of probe placement. To generate a depth-of-focus enhancing annular beam, we utilized a new fabrication method, whereby a hollow glass ferrule was angle-polished and gold-coated to produce an elongated annular reflector. We present validation data that verifies the preservation of linear scanning, despite the use of flexible materials. When utilized on excised, cultured mouse trachea, the probe acquired images of comparable quality to those obtained by a benchtop μOCT system.

High resolution micro-optical coherence tomography (µOCT) technology has been demonstrated to be useful for imaging respiratory epithelial functional microanatomy relevant to the study of pulmonary diseases such as cystic fibrosis and COPD. We previously reported the use of a benchtop μOCT imaging technology to image several relevant respiratory epithelial functional microanatomy at 40 fps and at lateral and axial resolutions of 2 and 1.3μm, respectively. We now present the development of a portable μOCT imaging system with comparable optical and imaging performance, which enables the μOCT technology to be translated to the clinic for in vivo imaging of human airways.

We have designed and fabricated a 4 mm diameter rigid endoscopic probe to obtain high resolution micro-optical coherence tomography (µOCT) images from the tracheal epithelium of living swine. Our common-path fiber-optic probe used gradient-index focusing optics, a selectively coated prism reflector to implement a circular-obscuration apodization for depth-of-focus enhancement, and a common-path reference arm and an ultra-broadbrand supercontinuum laser to achieve high axial resolution. Benchtop characterization demonstrated lateral and axial resolutions of 3.4 μm and 1.7 μm, respectively (in tissue). Mechanical standoff rails flanking the imaging window allowed the epithelial surface to be maintained in focus without disrupting mucus flow. During in vivo imaging, relative motion was mitigated by inflating an airway balloon to hold the standoff rails on the epithelium. Software implemented image stabilization was also implemented during post-processing. The resulting image sequences yielded co-registered quantitative outputs of airway surface liquid and periciliary liquid layer thicknesses, ciliary beat frequency, and mucociliary transport rate, metrics that directly indicate airway epithelial function that have dominated in vitro research in diseases such as cystic fibrosis, but have not been available in vivo.

Mucociliary clearance (MCC) plays a significant role in maintaining the health of human respiratory system by eliminating foreign particles trapped within mucus. Failure of this mechanism in diseases such as cystic fibrosis and chronic obstructive pulmonary disease (COPD) leads to airway blockage and lung infection, causing morbidity and mortality. The volume of airway mucus and the periciliary liquid encapsulating the cilia, in addition to ciliary beat frequency and velocity of mucociliary transport, are vital parameters of airway health. However, the diagnosis of disease pathogenesis and advances of novel therapeutics are hindered by the lack of tools for visualization of ciliary function in vivo. Our laboratory has previously developed a 1-µm resolution optical coherence tomography method, termed Micro-OCT, which is capable of visualizing mucociliary transport and quantitatively capturing epithelial functional metrics. We have also miniaturized Micro-OCT optics in a first-generation rigid 4mm Micro-OCT endoscope utilizing a common-path design and an apodizing prism configuration to produce an annular profile sample beam, and reported the first in vivo visualization of mucociliary transport in swine. We now demonstrate a flexible 2.5 mm Micro-OCT probe that can be inserted through the instrument channel of standard flexible bronchoscopes, allowing bronchoscopic navigation to smaller airways and greatly improving clinical utility. Longitudinal scanning over a field of view of more than 400 µm at a frame rate of 40 Hz was accomplished with a driveshaft transduced by a piezo-electric stack motor. We present characterization and imaging results from the flexible micro-OCT probe and progress towards clinical translation. The ability of the bronchoscope-compatible micro-OCT probe to image mucus clearance and epithelial function will enable studies of cystic fibrosis pathogenesis in small airways, provide diagnosis of mucociliary clearance disorders, and allow individual responses to treatments to be monitored.