In vivo assessment of inflammatory bowel disease in rats with ultrahigh-resolution colonoscopic OCT

A technology capable of high-resolution, label-free imaging of subtle pathology in vivo during colonoscopy is imperative for the early detection of disease and the performance of accurate biopsies. While colonoscopic OCT has been developed to visualize colonic microstructures beyond the mucosal surface, its clinical potential remains limited by sub-optimal resolution (∼6.5 µm in tissue), inadequate imaging contrast, and a lack of high-resolution OCT criteria for lesion detection. In this study, we developed an ultrahigh-resolution (UHR) colonoscopic OCT and evaluated its ability to volumetrically visualize and identify the pathological features of inflammatory bowel disease (IBD) in a rat model. Owing to its improved resolution (∼1.7 µm in tissue) and enhanced contrast, UHR colonoscopic OCT can accurately delineate fine colonic microstructures and identify the pathophysiological characteristics of IBD in vivo. By using a quantitative optical attenuation map, UHR colonoscopic OCT is able to differentiate diseased tissue (such as crypt distortion and microabscess) from normal colonic mucosa over a large field of view in vivo. Our results suggest the clinical potential of UHR colonoscopic OCT for in vivo assessment of IBD pathology.     

Corneal imaging with blue-light optical coherence microscopy

Corneal imaging is important for the diagnostic and therapeutic evaluation of many eye diseases. Optical coherence tomography (OCT) is extensively used in ocular imaging due to its non-invasive and high-resolution volumetric imaging characteristics. Optical coherence microscopy (OCM) is a technical variation of OCT that can image the cornea with cellular resolution. Here, we demonstrate a blue-light OCM as a low-cost and easily reproducible system to visualize corneal cellular structures such as epithelial cells, endothelial cells, keratocytes, and collagen bundles within stromal lamellae. Our blue-light OCM system achieved an axial resolution of 12 µm in tissue over a 1.2 mm imaging depth, and a lateral resolution of 1.6 µm over a field of view of 750 µm × 750 µm.     

Miniature real-time intraoperative forward-imaging optical coherence tomography probe

Optical coherence tomography (OCT) has a tremendous global impact upon the ability to diagnose, treat, and monitor eye diseases. A miniature 25-gauge forward-imaging OCT probe with a disposable tip was developed for real-time intraoperative ocular imaging of posterior pole and peripheral structures to improve vitreoretinal surgery. The scanning range was 2 mm when the probe tip was held 3-4 mm from the tissue surface. The axial resolution was 4-6 µm and the lateral resolution was 25-35 µm. The probe was used to image cellophane tape and multiple ocular structures.OCIS codes: (170.4500) Optical coherence tomography, (120.3890) Medical optics instrumentation     

Microscopic optical coherence tomography (mOCT) at 600 kHz for 4D volumetric imaging and dynamic contrast

Volumetric imaging of dynamic processes with microscopic resolution holds a huge potential in biomedical research and clinical diagnosis. Using supercontinuum light sources and high numerical aperture (NA) objectives, optical coherence tomography (OCT) achieves microscopic resolution and is well suited for imaging cellular and subcellular structures of biological tissues. Currently, the imaging speed of microscopic OCT (mOCT) is limited by the line-scan rate of the spectrometer camera and ranges from 30 to 250 kHz. This is not fast enough for volumetric imaging of dynamic processes in vivo and limits endoscopic application. Using a novel CMOS camera, we demonstrate fast 3-dimensional OCT imaging with 600,000 A-scans/s at 1.8 µm axial and 1.1 µm lateral resolution. The improved speed is used for imaging of ciliary motion and particle transport in ex vivo mouse trachea. Furthermore, we demonstrate dynamic contrast OCT by evaluating the recorded volumes rather than en face planes or B-scans. High-speed volumetric mOCT will enable the correction of global tissue motion and is a prerequisite for applying dynamic contrast mOCT in vivo. With further increase in imaging speed and integration in flexible endoscopes, volumetric mOCT may be used to complement or partly replace biopsies.     

Ultrahigh speed endoscopic optical coherence tomography for gastroenterology

We describe an ultrahigh speed endoscopic swept source optical coherence tomography (OCT) system for clinical gastroenterology using a vertical-cavity surface-emitting laser (VCSEL) and micromotor imaging catheter. The system had a 600 kHz axial scan rate and 8 µm axial resolution in tissue. Imaging was performed with a 3.2 mm diameter imaging catheter at 400 frames per second with a 12 µm spot size. Three-dimensional OCT (3D-OCT) imaging was performed in patients with a cross section of pathologies undergoing upper and lower endoscopy. The use of distally actuated imaging catheters enabled OCT imaging with more flexibility, such as volumetric imaging in the small intestine and the assessment of hiatal hernia using retroflex imaging. The high rotational scanning stability of the micromotor enabled 3D volumetric imaging with micron scale volumetric accuracy for both en face OCT and cross-sectional imaging, as well as OCT angiography (OCTA) for 3D visualization of subsurface microvasculature. The ability to perform both structural and functional 3D OCT imaging in the GI tract with microscopic accuracy should enable a wide range of studies and enhance the sensitivity and specificity of OCT for detecting pathology.OCIS codes: (110.2350) Fiber optics imaging, (120.3890) Medical optics instrumentation, (120.5800) Scanners, (110.6880) Three-dimensional image acquisition, (140.7260) Vertical cavity surface emitting lasers, (170.2150) Endoscopic imaging, (170.2680) Gastrointestinal, (170.3880) Medical and biological imaging, (170.4500) Optical coherence tomography     

Quantitative spectroscopic comparison of the optical properties of mouse cochlea microstructures using optical coherence tomography at 1.06 µm and 1.3 µm wavelengths

Currently, the cochlear implantation procedure mainly relies on using a hand lens or surgical microscope, where the success rate and surgery time strongly depend on the surgeon’s experience. Therefore, a real-time image guidance tool may facilitate the implantation procedure. In this study, we performed a systematic and quantitative analysis on the optical characterization of ex vivo mouse cochlear samples using two swept-source optical coherence tomography (OCT) systems operating at the 1.06-µm and 1.3-µm wavelengths. The analysis results demonstrated that the 1.06-µm OCT imaging system performed better than the 1.3-µm OCT imaging system in terms of the image contrast between the cochlear conduits and the neighboring cochlear bony wall structure. However, the 1.3-µm OCT imaging system allowed for greater imaging depth of the cochlear samples because of decreased tissue scattering. In addition, we have investigated the feasibility of identifying the electrode of the cochlear implant within the ex vivo cochlear sample with the 1.06-µm OCT imaging. The study results demonstrated the potential of developing an image guidance tool for the cochlea implantation procedure as well as other otorhinolaryngology applications.     

Broadband miniature optical ultrasound probe for high resolution vascular tissue imaging

An all-optical ultrasound probe for vascular tissue imaging was developed. Ultrasound was generated by pulsed laser illumination of a functionalized carbon nanotube composite coating on the end face of an optical fiber. Ultrasound was detected with a Fabry-Pérot (FP) cavity on the end face of an adjacent optical fiber. The probe diameter was < 0.84 mm and had an ultrasound bandwidth of ~20 MHz. The probe was translated across the tissue sample to create a virtual linear array of ultrasound transmit/receive elements. At a depth of 3.5 mm, the axial resolution was 64 µm and the lateral resolution was 88 µm, as measured with a carbon fiber target. Vascular tissues from swine were imaged ex vivo and good correspondence to histology was observed.OCIS codes: (110.5125) Photoacoustics, (110.2350) Fiber optics imaging, (060.2380) Fiber optics sources and detectors, (170.7170) Ultrasound, (170.0110) Imaging systems     

Piezoelectric-transducer-based miniature catheter for ultrahigh-speed endoscopic optical coherence tomography

We developed a piezoelectric-transducer- (PZT) based miniature catheter with an outer diameter of 3.5 mm for ultrahigh-speed endoscopic optical coherence tomography (OCT). A miniaturized PZT bender actuates a fiber and the beam is scanned through a GRIN lens and micro-prism to provide high-speed, side-viewing capability. The probe optics can be pulled back over a long distance to acquire three-dimensional (3D) data sets covering a large area. Imaging is performed with 11 μm axial resolution in air (8 μm in tissue) and 20 μm transverse resolution, at 960 frames per second with a Fourier domain mode-locked laser operating at 480 kHz axial scan rate. Using a high-speed data acquisition system, endoscopic OCT imaging of the rabbit esophagus and colon in vivo and human colon specimens ex vivo is demonstrated.     

In vivo three- and four-photon fluorescence microscopy using a 1.8 µm femtosecond fiber laser system

Multiphoton microscopy has enabled us to image cellular dynamics in vivo. However, the excitation wavelength for imaging with commercially available lasers is mostly limited between 0.65–1.04 µm. Here we develop a femtosecond fiber laser system that produces ∼150 fs pulses at 1.8 µm. Our system starts from an erbium-doped silica fiber laser, and its wavelength is converted to 1.8 µm using a Raman shift fiber. The 1.8 µm pulses are amplified with a two-stage Tm:ZBLAN fiber amplifier. The final pulse energy is ∼1 µJ, sufficient for in vivo imaging. We successfully observe TurboFP635-expressing cortical neurons at a depth of 0.7 mm from the brain surface by three-photon excitation and Clover-expressing astrocytes at a depth of 0.15 mm by four-photon excitation.     

Ultra-widefield retinal MHz-OCT imaging with up to 100 degrees viewing angle

We evaluate strategies to maximize the field of view (FOV) of in vivo retinal OCT imaging of human eyes. Three imaging modes are tested: Single volume imaging with 85° FOV as well as with 100° and stitching of five 60° images to a 100° mosaic (measured from the nodal point). We employ a MHz-OCT system based on a 1060nm Fourier domain mode locked (FDML) laser with a depth scan rate of 1.68MHz. The high speed is essential for dense isotropic sampling of the large areas. Challenges caused by the wide FOV are discussed and solutions to most issues are presented. Detailed information on the design and characterization of our sample arm optics is given. We investigate the origin of an angle dependent signal fall-off which we observe towards larger imaging angles. It is present in our 85° and 100° single volume images, but not in the mosaic. Our results suggest that 100° FOV OCT is possible with current swept source OCT technology.OCIS codes: (170.4500) Optical coherence tomography, (170.3880) Medical and biological imaging, (170.4460) Ophthalmic optics and devices, (120.3890) Medical optics instrumentation, (140.3510) Lasers, fiber     

Adaptive optics scanning laser ophthalmoscopy and optical coherence tomography (AO-SLO-OCT) system for in vivo mouse retina imaging

Optical coherence tomography (OCT) and scanning laser ophthalmoscopy (SLO) are imaging technologies invented in the 1980s that have revolutionized the field of in vivo retinal diagnostics and are now commonly used in ophthalmology clinics as well as in vision science research. Adaptive optics (AO) technology enables high-fidelity correction of ocular aberrations, resulting in improved resolution and sensitivity for both SLO and OCT systems. The potential of gathering multi-modal cellular-resolution information in a single instrument is of great interest to the ophthalmic imaging community. Although similar instruments have been developed for imaging the human retina, developing such a system for mice will benefit basic science research and should help with further dissemination of AO technology. Here, we present our work integrating OCT into an existing mouse retinal AO-SLO system, resulting in a multi-modal AO-enhanced imaging system of the living mouse eye. The new system allows either independent or simultaneous data acquisition of AO-SLO and AO-OCT, depending on the requirements of specific scientific experiments. The system allows a data acquisition speed of 200 kHz A-scans/pixel rate for OCT and SLO, respectively. It offers ∼6 µm axial resolution for AO-OCT and a ∼1 µm lateral resolution for AO-SLO-OCT imaging.     

Deep-learning-aided forward optical coherence tomography endoscope for percutaneous nephrostomy guidance

Percutaneous renal access is the critical initial step in many medical settings. In order to obtain the best surgical outcome with minimum patient morbidity, an improved method for access to the renal calyx is needed. In our study, we built a forward-view optical coherence tomography (OCT) endoscopic system for percutaneous nephrostomy (PCN) guidance. Porcine kidneys were imaged in our experiment to demonstrate the feasibility of the imaging system. Three tissue types of porcine kidneys (renal cortex, medulla, and calyx) can be clearly distinguished due to the morphological and tissue differences from the OCT endoscopic images. To further improve the guidance efficacy and reduce the learning burden of the clinical doctors, a deep-learning-based computer aided diagnosis platform was developed to automatically classify the OCT images by the renal tissue types. Convolutional neural networks (CNN) were developed with labeled OCT images based on the ResNet34, MobileNetv2 and ResNet50 architectures. Nested cross-validation and testing was used to benchmark the classification performance with uncertainty quantification over 10 kidneys, which demonstrated robust performance over substantial biological variability among kidneys. ResNet50-based CNN models achieved an average classification accuracy of 82.6%±3.0%. The classification precisions were 79%±4% for cortex, 85%±6% for medulla, and 91%±5% for calyx and the classification recalls were 68%±11% for cortex, 91%±4% for medulla, and 89%±3% for calyx. Interpretation of the CNN predictions showed the discriminative characteristics in the OCT images of the three renal tissue types. The results validated the technical feasibility of using this novel imaging platform to automatically recognize the images of renal tissue structures ahead of the PCN needle in PCN surgery.     

A high-efficiency fiber-based imaging system for co-registered autofluorescence and optical coherence tomography

We present a power-efficient fiber-based imaging system capable of co-registered autofluorescence imaging and optical coherence tomography (AF/OCT). The system employs a custom fiber optic rotary joint (FORJ) with an embedded dichroic mirror to efficiently combine the OCT and AF pathways. This three-port wavelength multiplexing FORJ setup has a throughput of more than 83% for collected AF emission, significantly more efficient compared to previously reported fiber-based methods. A custom 900 µm diameter catheter ‒ consisting of a rotating lens assembly, double-clad fiber (DCF), and torque cable in a stationary plastic tube ‒ was fabricated to allow AF/OCT imaging of small airways in vivo. We demonstrate the performance of this system ex vivo in resected porcine airway specimens and in vivo in human on fingers, in the oral cavity, and in peripheral airways.OCIS codes: (110.0110) Imaging systems, (110.2350) Fiber optics imaging, (110.4500) Optical coherence tomography, (170.2520) Fluorescence microscopy, (170.3890) Medical optics instrumentation     

Miniature structured illumination microscope for in vivo 3D imaging of brain structures with optical sectioning

We present a high-resolution miniature, light-weight fluorescence microscope with electrowetting lens and onboard CMOS for high resolution volumetric imaging and structured illumination for rejection of out-of-focus and scattered light. The miniature microscope (SIMscope3D) delivers structured light using a coherent fiber bundle to obtain optical sectioning with an axial resolution of 18 µm. Volumetric imaging of eGFP labeled cells in fixed mouse brain tissue at depths up to 260 µm is demonstrated. The functionality of SIMscope3D to provide background free 3D imaging is shown by recording time series of microglia dynamics in awake mice at depths up to 120 µm in the brain.     

Ultra-thin and flexible endoscopy probe for optical coherence tomography based on stepwise transitional core fiber

We present an ultra-thin fiber-body endoscopy probe for optical coherence tomography (OCT) which is based on a stepwise transitional core (STC) fiber. In a minimalistic design, our probe was made of spliced specialty fibers that could be directly used for beam probing optics without using a lens. In our probe, the OCT light delivered through a single-mode fiber was efficiently expanded to a large mode field of 24 μm diameter for a low beam divergence. The size of our probe was 85 μm in the probe’s diameter while operated in a 160-μm thick protective tubing. Through theoretical and experimental analyses, our probe was found to exhibit various attractive features in terms of compactness, flexibility and reliability along with its excellent fabrication simplicity.OCIS codes: (110.4500) Optical coherence tomography, (170.2150) Endoscopic imaging, (170.3890) Medical optics instrumentation, (060.2350) Fiber optics imaging     

Motion-compensated hand-held common-path Fourier-domain optical coherence tomography probe for image-guided intervention

A motion-compensated, hand-held, common-path, Fourier-domain optical coherence tomography imaging probe has been developed for image-guided intervention during microsurgery. A hand-held prototype instrument was achieved by integrating an imaging fiber probe inside a stainless steel needle and attached to the ceramic shaft of a piezoelectric motor housed in an aluminum handle. The fiber probe obtains A-scan images. The distance information was extracted from the A-scans to track the sample surface distance and a fixed distance was maintained by a feedback motor control which effectively compensated hand tremor and target movements in the axial direction. Real-time data acquisition, processing, motion compensation, and image visualization and saving were implemented on a custom CPU-GPU hybrid architecture. We performed 10× zero padding to the raw spectrum to obtain 0.16 µm position accuracy with a compensation rate of 460 Hz. The root-mean-square error of hand-held distance variation from target position was measured to be 2.93 µm. We used a cross-correlation maximization-based shift correction algorithm for topology correction. To validate the system, we performed free-hand OCT M-scan imaging using various samples.OCIS codes: (100.2000) Digital image processing, (110.4500) Optical coherence tomography, (170.3890) Medical optics instrumentation     

Automatic identification of the temporal retinal nerve fiber raphe from macular cube data

We evaluated several approaches for automatic location of the temporal nerve fiber raphe from standard macular cubes acquired on a Heidelberg Spectralis OCT. Macular cubes with B-scan separation of 96–122 µm were acquired from 15 healthy participants, and “high density” cubes with scan separation of 11 µm were acquired from the same eyes. These latter scans were assigned to experienced graders for subjective location of the raphe, providing the ground truth by which to compare methods operating on the lower density data. A variety of OCT scan parameters and image processing strategies were trialed. Vertically oriented scans, purposeful misalignment of the pupil to avoid reflective artifacts, and the use of intensity as opposed to thickness of the nerve fiber layer were all critical to minimize error. The best performing approach “cFan” involved projection of a fan of lines from each of several locations across the foveal pit; in each fan the line of least average intensity was identified. The centroid of the crossing points of these lines provided the raphe orientation with an average error of 1.5° (max = 4.1°) relative to the human graders. The disc-fovea-raphe angle was 172.4 ± 2.3° (range = 168.5–176.2°), which agrees well with other published estimates.OCIS codes: (170.4500) Optical coherence tomography, (100.0100) Image processing, (100.3008) Image recognition, algorithms and filters, (170.4580) Optical diagnostics for medicine     

Ultrahigh resolution spectral-domain optical coherence tomography using the 1000–1600 nm spectral band

Ultrahigh resolution optical coherence tomography (UHR-OCT) can image microscopic features that are not visible with the standard OCT resolution of 5-15 µm. In previous studies, high-speed UHR-OCT has been accomplished within the visible (VIS) and near-infrared (NIR-I) spectral ranges, specifically within 550-950 nm. Here, we present a spectral domain UHR-OCT system operating in a short-wavelength infrared (SWIR) range from 1000 to 1600 nm using a supercontinuum light source and an InGaAs-based spectrometer. We obtained an axial resolution of 2.6 µm in air, the highest ever recorded in the SWIR window to our knowledge, with deeper penetration into tissues than VIS or NIR-I light. We demonstrate imaging of conduction fibers of the left bundle branch in freshly excised porcine hearts. These results suggest a potential for deep-penetration, ultrahigh resolution OCT in intraoperative applications.     

Multimodal nonlinear endo-microscopy probe design for high resolution,label-free intraoperative imaging

We present a portable, multimodal, nonlinear endo-microscopy probe designed for intraoperative oncological imaging. Application of a four-wave mixing noise suppression scheme using dual wavelength wave plates (DWW) and a polarization-maintaining fiber improves tissue signal collection efficiency, allowing for miniaturization. The probe, with a small 14 mm transversal diameter, includes a customized miniaturized two-axis MEMS (micro-electromechanical system) raster scanning mirror and micro-optics with an illumination laser delivered by a polarization-maintaining fiber. The probe can potentially be integrated into the arms of a surgical robot, such as da Vinci robotic surgery system, due to its minimal cross sectional area. It has the ability to incorporate multiple imaging modalities including CARS (coherent anti-Stokes Raman scattering), SHG (second harmonic generation), and TPEF (two-photon excited fluorescence) in order to allow the surgeon to locate tumor cells within the context of normal stromal tissue. The resolution of the endo-microscope is experimentally determined to be 0.78 µm, a high level of accuracy for such a compact probe setup. The expected resolution of the as-built multimodal, nonlinear, endo-microscopy probe is 1 µm based on the calculation tolerance allocation using Monte-Carlo simulation. The reported probe is intended for use in laparoscopic or radical prostatectomy, including detection of tumor margins and avoidance of nerve impairment during surgery.OCIS codes: (110.0110) Imaging systems, (220.0220) Optical design and fabrication     

Characterization of eosinophilic esophagitis murine models using optical coherence tomography

Pre-clinical studies using murine models are critical for understanding the pathophysiological mechanisms underlying immune-mediated disorders such as Eosinophilic esophagitis (EoE). In this study, an optical coherence tomography (OCT) system capable of providing three-dimensional images with axial and transverse resolutions of 5 µm and 10 µm, respectively, was utilized to obtain esophageal images from a murine model of EoE-like disease ex vivo. Structural changes in the esophagus of wild-type (Tslpr^+/+) and mutant (Tslpr^−/−) mice with EoE-like disease were quantitatively evaluated and food impaction sites in the esophagus of diseased mice were monitored using OCT. Here, the capability of OCT as a label-free imaging tool devoid of tissue-processing artifacts to effectively characterize murine EoE-like disease models has been demonstrated.OCIS codes: (110.4500) Optical coherence tomography, (110.6880) Three-dimensional image acquisition, (170.0110) Imaging systems, (170.2680) Gastrointestinal, (170.3880) Medical and biological imaging, (170.6935) Tissue characterization