Handheld common-path swept-source optical coherence tomography angiography.

This study develops a handheld optical coherence tomography angiography (OCTA) system that uses a high-speed (200 kHz) swept laser with a dual-reference common-path configuration for stable and fast imaging. The common-path design automatically avoids polarization and dispersion mismatches by using one circulator as the primary system element, ensuring a cost-effective and compact design for handheld probe use. With its stable envelope (i.e., sub-µm shifts) and phase variation (corresponding to nm changes in axial displacement), the minimum detectable flow velocity is ∼ 0.08 mm/s in our experiment, which gives the common-path setup a high potential for application in a handheld OCTA system for clinical skin screening. In vivo skin structures and microvasculature networks on the dorsum of the hand and cheek of a healthy human are imaged successfully.

Fiber-Needle Swept-Source Optical Coherence Tomography for the Real-Time Visualization of the Transversus Abdominis Plane Block Procedure in a Swine Model.

BACKGROUND: Fascia blocks (eg, the transversus abdominis plane [TAP] block) target the intermuscular fascia layers. Ultrasound techniques have allowed peripheral blocks to be performed with accuracy and safety, however, with limitations. Optical coherence tomography (OCT) is based on low-coherence interferometry. In this study, we examined the ability of OCT to identify the TAP. METHODS: A swept-source OCT probe was placed in a 17-gauge needle to obtain imaging. The needle was inserted within 2 different angle ranges (0°-30° and 30°-60°) on a slice of pork belly to assess imaging characteristics. A series of real-time OCT imaging of the muscle, fascia, and interfascial space was obtained. The tissue location of the needle tip was identified using near-infrared (NIR) imaging. In vivo OCT imaging was further done on 3 female 6-month-old native Chinese Landrance Duroc pigs. Real-time images of tissue layers were obtained with needle insertion. Ultrasound imaging of the OCT needle probe was also performed at the same time for needle trajectory guidance. After imaging, the OCT probe was removed, and 5 mL of normal saline was injected via the needle to confirm correct fascia plane identification. RESULTS: In and ex vivo studies showed clear visual distinction of muscle, fascia, and interfascial layer with OCT, with limitations. Independent validation of OCT criteria for the muscle/fascia differentiation by 20 OCT readers for the in vivo data demonstrated the sensitivity = 0.91, specificity = 0.90, and accuracy = 0.89. Although the angle of needle entry affected the depth of OCT penetration in the muscle, the attenuation coefficient values of the fascia and muscle tissue were statistically different (P < .001) and with high area under the receiver operating characteristics (ROC) curve (AUC) (AUC = 0.93 in 0°-30° and AUC = 1 in 30°-60°) for fascia identification. CONCLUSIONS: This study introduced a novel needle imaging probe method to identify the transversus abdominis fascia plane in real-time. Quantitative calculation of the attenuation coefficients can further aid objective identification by providing direct confirmation of the tip position, increasing the first-pass success rate, and decreasing the need for needle repositioning. Combining OCT and ultrasound may improve the accuracy of anesthetics placement.

Fiber-needle swept-source optical coherence tomography system for the identification of the epidural space in piglets.

BACKGROUND: Epidural needle insertion is traditionally a blind technique whose success depends on the experience of the operator. The authors describe a novel method using a fiber-needle-based swept-source optical coherence tomography (SSOCT) to identify epidural space. METHODS: An optical fiber probe was placed into a hollow 18-gauge Tuohy needle. It was then inserted by an experienced anesthesiologist to continuously construct a series of two-dimensional SSOCT images by mechanically rotating the optical probe. To quantify this observation, both the average SSOCT signal intensities and their diagnostic potentials were assessed. The insertions were performed three times into both the lumbar and thoracic regions of five pigs using a paramedian approach. RESULTS: A side-looking SSOCT is constructed to create a visual image of the underlying structures. The image criteria for the identification of the epidural space from the outside region were generated by the analysis of a training set (n = 100) of ex vivo data. The SSOCT image criteria for in vivo epidural space identification are high sensitivity (0.867 to 0.965) and high specificity (0.838 to 0.935). The mean value of the average signal intensities exhibits statistically significant differences (P < 0.01) and a high discriminatory capacity (area under curve = 0.88) between the epidural space and the outside tissues. CONCLUSIONS: This is the first study to introduce a SSOCT fiber probe embedded in a standard epidural needle. The authors anticipate that this technique will reduce the occurrence of failed epidural blocks and other complications such as dural punctures.

Miniature endoscopic optical coherence tomography for calculus detection.

The effective treatment of periodontitis involves the detection and removal of subgingival dental calculus. However, subgingival calculus is more difficult to detect than supragingival calculus because it is firmly attached to root surfaces within periodontal pockets. To achieve a smooth root surface, clinicians often remove excessive amounts of root structure because of decreased visibility. In addition, enamel pearl, a rare type of ectopic enamel formation on the root surface, can easily be confused with dental calculus in the subgingival environment. In this study, we developed a fiber-probe swept-source optical coherence tomography (SSOCT) technique and combined it with the quantitative measurement of an optical parameter [standard deviation (SD) of the optical coherence tomography (OCT) intensity] to differentiate subgingival calculus from sound enamel, including enamel pearl. Two-dimensional circumferential images were constructed by rotating the miniprobe (0.9 mm diameter) while acquiring image lines, and the adjacent lines in each rotation were stacked to generate a three-dimensional volume. In OCT images, compared to sound enamel and enamel pearls, dental calculus showed significant differences (P<0.001) in SD values. Finally, the receiver operating characteristic curve had a high capacity (area under the curve=0.934) for discriminating between healthy regions (including enamel pearl) and dental calculus.

Balanced detection for spectral domain optical coherence tomography.

The use and advantages of applying balanced-detection (BD) operation method to high speed spectral domain optical coherence tomography (SDOCT) are presented in this study, which we believe is the first such demonstration. Compared to conventional SDOCT, BD-SDOCT provides two unique advantages. First, the method can suppress background noise and autocorrelation artifacts in biological tissues. Second, it is a power-efficient method which is particularly helpful for high speed SDOCT to eliminate random intensity noise and to achieve shot noise limited detection. This performance allows in vivo three-dimensional tissue visualization with high imaging quality and high speed.

Quantification of microvascular change of retinal degeneration in Royal College of Surgeons rats using high-resolution spectral domain optical coherence tomography angiography.

Significance: For research on retinitis pigmentosa in humans, the Royal College of Surgeons (RCS) rat is commonly used as the primary animal model since the disease process is similar. Therefore, it is necessary to understand how the disease develops and determine whether the treatment is effective. Aim: In this study, structural and microvascular change of retinal degeneration in RCS rats was assessed non-invasively on specific dates over 3.5 months. Approach: Using a high-resolution spectral domain (SD) optical coherence tomography angiography (OCTA), the retinal degeneration in RCS rats, from day 14 until day 126, was qualitatively and quantitatively analyzed. Results: Aside from the thinning of the retina thickness starting from 2 weeks of age, blood vessels in the deep layer of the retina also began to degenerate at about 4 weeks of age. Hole structures appeared at the inner nuclear layer and the inner plexiform layer by the age of 10 weeks. Observations of abnormal angiogenesis in the choroid began by 12 weeks of age. Conclusions: We conducted a longitudinal study of retina degeneration structure and vascular changes in an RCS rat model using a supercontinuum laser based high-resolution SD-OCTA. Combined with OCTA, OCT leads to a better understanding of photoreceptor pathology as retinal degeneration by identifying tissue and vessel loss.

Inhibition of oxidative stress-induced epithelial-mesenchymal transition in retinal pigment epithelial cells of age-related macular degeneration model by suppressing ERK activation.

INTRODUCTION: Epithelial-mesenchymal transition (EMT) of retinal pigment epithelial (RPE) cells is related to the pathogenesis of various retinopathies including age-related macular degeneration (AMD). Oxidative stress is the major factor that induces degeneration of RPE cells associated with the etiology of AMD. OBJECTIVES: Sodium iodate (NaIO3) generates intracellular reactive oxygen species (ROS) and is widely used to establish a model of AMD due to the selective induction of retinal degeneration. This study was performed to clarify the effects of multiple NaIO3-stimulated signaling pathways on EMT in RPE cells. METHODS: The EMT characteristics in NaIO3-treated human ARPE-19 cells and RPE cells of the mouse eyes were analyzed. Multiple oxidative stress-induced modulators were investigated and the effects of pre-treatment with Ca2+ chelator, extracellular signal-related kinase (ERK) inhibitor, or epidermal growth factor receptor (EGFR) inhibitor on NaIO3-induced EMT were determined. The efficacy of post-treatment with ERK inhibitor on the regulation of NaIO3-induced signaling pathways was dissected and its role in retinal thickness and morphology was evaluated by using histological cross-sections and spectral domain optical coherence tomography. RESULTS: We found that NaIO3 induced EMT in ARPE-19 cells and in RPE cells of the mouse eyes. The intracellular ROS, Ca2+, endoplasmic reticulum (ER) stress marker, phospho-ERK, and phospho-EGFR were increased in NaIO3-stimulated cells. Our results showed that pre-treatment with Ca2+ chelator, ERK inhibitor, or EGFR inhibitor decreased NaIO3-induced EMT, interestingly, the inhibition of ERK displayed the most prominent effect. Furthermore, post-treatment with FR180204, a specific ERK inhibitor, reduced intracellular ROS and Ca2+ levels, downregulated phospho-EGFR and ER stress marker, attenuated EMT of RPE cells, and prevented structural disorder of the retina induced by NaIO3. CONCLUSIONS: ERK is a crucial regulator of multiple NaIO3-induced signaling pathways that coordinate EMT program in RPE cells. Inhibition of ERK may be a potential therapeutic strategy for the treatment of AMD.

Balanced detection spectral domain optical coherence tomography with a multiline single camera for signal-to-noise ratio enhancement.

In this study, the use and advantages of balanced detection (BD) in spectral domain optical coherence tomography (SD-OCT) are demonstrated. A-scans are calculated as a combination of two phase-opposed interferometric spectra acquired simultaneously by using a multiline single camera spectrometer. Not only does this system suppress artifacts due to autocorrelation, but also the signal of interest is increased by a factor of 2 as experimentally verified. Our BD-based SD-OCT gives a signal-to-noise ratio improvement of 8-14 dB for the peak within 1 mm compared to standard SD-OCT using a single detection scheme. This method is validated by experimental measurement of a glass plate.

Needle-Probe Optical Coherence Tomography for Real-Time Visualization of Veress Peritoneal Needle Placement in a Porcine Model: A New Safety Concept for Pneumoperitoneum Establishment in Laparoscopic Surgery.

The safe establishment of pneumoperitoneum is a critical step in all laparoscopic surgeries. A closed pneumoperitoneum is usually obtained by inserting a Veress needle into the peritoneal cavity. However, there is no definite measure to visually confirm the position of the Veress needle tip inside the peritoneal cavity. This study aimed to describe a method of real-time visual detection of peritoneal placement of the Veress needle using an incorporated optical coherence tomography (OCT) probe in a porcine model. A 14-gauge Veress needle was incorporated with a miniature fiber probe to puncture the piglet's abdominal wall into the peritoneal cavity. A total of 80 peritoneal punctures were attempted in four piglets. For each puncture, continuous two-dimensional OCT images of the abdominal wall were acquired for real-time visual detection of the needle placement into the peritoneal cavity. Characteristic OCT image patterns could be observed during the puncturing process, especially a deep V-shaped concave pattern before the peritoneum puncture, which was a crucial feature. A statistical difference in the OCT signal standard deviation value also indicated the differentiability of images between the peritoneum and extra-peritoneal tissue layers. A success rate of 97.5% could be achieved with the guidance of the OCT images. OCT images translate the blind closed technique of peritoneal access into a visualized procedure, thus improving peritoneal access safety.

Human Umbilical Mesenchymal Stem Cell Xenografts Repair UV-Induced Photokeratitis in a Rat Model.

Most patients with a corneal injury are administered anti-inflammatory medications and antibiotics, but no other treatments are currently available. Thus, the corneal injury healing is unsatisfactory, affects the vision, and has a risk of blindness in severe cases. Human umbilical mesenchymal stem cells exhibit pluripotent and anti-inflammatory properties and do not cause immunological rejection in the host. Rats were irradiated with type B ultraviolet (UVB) light to generate a stable animal model of photokeratitis. After irradiation-induced photokeratitis, human umbilical mesenchymal stem cells were implanted into the subconjunctival space of the lateral sclera, and the changes in the corneal pathology were evaluated. Three weeks after implantation, many mesenchymal stem cells were visible in the subconjunctival space. These mesenchymal stem cells effectively reduced the extent of injury to the adjacent corneal tissue. They accelerated the epithelial layer repair, reduced the inflammatory response and neovascularization, and improved the disorganization of collagen and fibronectin in the corneal stroma caused by the injury. In conclusion, xenografted human umbilical mesenchymal stem cells can survive in rat eye tissues for a long time, effectively support the structural integrity of injured corneal tissues, restore corneal permeability, and reduce abnormal neovascularization. This study provides a new approach to the treatment of photokeratitis.

En-face polarization-sensitive optical coherence tomography to characterize early-stage esophageal cancer and determine tumor margin.

Current imaging tools are insufficiently sensitive to the early diagnosis of esophageal squamous cell carcinoma (ESCC). The application of polarization-sensitive optical coherence tomography (PS-OCT) to detect tumor-stroma interaction is an interesting issue in cancer diagnosis. In this translational study, we found that en-face PS-OCT effectively characterizes protruding, flat, and depressive type ESCC regardless of animal or human specimens. In addition, the tumor contour and margin could also be drawn and determined on a broad en-face view. The determined tumor margin could be in the proximity of 2 mm to the actual tumor margin, which was proved directly using histology.

Inhibition of DUSP6 Activates Autophagy and Rescues the Retinal Pigment Epithelium in Sodium Iodate-Induced Retinal Degeneration Models In Vivo and In Vitro.

Autophagy plays a protective role in the retinal pigment epithelium (RPE) by eliminating damaged organelles in response to reactive oxygen species (ROS). Dual-specificity protein phosphatase 6 (DUSP6), which belongs to the DUSP subfamily, works as a negative-feedback regulator of the extracellular signal-regulated kinase (ERK) pathway. However, the complex interplay between DUSP6 and autophagy induced by ROS in RPE is yet to be investigated. To investigate the relationship between DUSP6 and autophagy, we exposed the ARPE-19 cell line and C57BL/6N mice to sodium iodate (NaIO3) as an oxidative stress inducer. Our data showed that the inhibition of DUSP6 activity promotes autophagy flux through the ERK pathway via the upregulation of immunoblotting expression in ARPE-19 cells. Live imaging showed a significant increase in autophagic flux activities, which suggested the restoration autophagy after treatment with the DUSP6 inhibitor. Furthermore, the mouse RPE layer exhibited an irregular structure and abnormal deposits following NaIO3 injection. The retina layer was recovered after being treated with DUSP6 inhibitor; this suggests that DUSP6 inhibitor can rescue retinal damage by restoring the mouse retina's autophagy flux. This study suggests that the upregulation of DUSP6 can cause autophagy flux malfunctions in the RPE. The DUSP6 inhibitor can restore autophagy induction, which may serve as a potential therapeutic approach for retinal degeneration disease.

In Vivo Longitudinal Tracking of Lymphangiogenesis and Angiogenesis in Cutaneous Melanoma Mouse Model Using Multifunctional Optical Coherence Tomography.

Melanoma is a high-risk skin cancer because it tends to metastasize early and ultimately leads to death. In this study, we introduced a noninvasive multifunctional optical coherence tomography (MFOCT) for the early detection of premetastatic pathogenesis in cutaneous melanoma by label-free imaging of microstructures (i.e., providing the thickness and the scattering information) and microcirculation (i.e., providing depth-resolved angiography and lymphangiography). Using MFOCT-based approaches, we presented an in vivo longitudinal observation of the tumor microenvironment in Braf V600E/V600E ;Pten -/- mice with inducible melanoma monitored for 42 days. Quantitative analysis of MFOCT images identified an increased number of lymphatic and vascular vessels during tumor progression and faster lymphangiogenesis (beginning on day 21) than angiogenesis (beginning on day 28) in the melanoma microenvironment. We further observed lymphatic vessel enlargement from the first week of melanoma development, implying tumor cells interacting with the vessels and increased likelihood of metastasis. MFOCT identified cutaneous melanoma‒associated angiogenesis and lymphangiogenesis before the possible visual perception of the tumor (≥42 days) and before metastasis could be diagnosed using micropositron emission tomography (35 days). Thus, the proposed quantitative analysis using MFOCT has the potential for early detection of cutaneous melanoma progression or prediction of metastatic melanoma in a mouse model. However, retrospective and extensive experiments still need to be performed in the future to confirm the value of MFOCT in clinical application.

Cell parameter measurement of a twisted nematic liquid crystal device using interferometric polarimeter under normal incidence.

Five cell parameters of a twisted nematic liquid crystal device (TNLCD), namely, cell gap, pretilt angle, twisted angle, rubbing angle, and phase retardation are precisely measured by the developed amplitude-sensitive heterodyne polarimeter (ASHP) simultaneously integrated with Yeh and Gu's transfer matrix and Lien's transfer matrix. This proposed method can characterize the five cell parameters under the arrangement of a single wavelength at normal incidence. In contrast to the conventional methods on cell parameter detection either by adopting a multiple wavelength laser beam at normal incidence or by using a single wavelength laser beam under oblique incident to TNLCD, this method presents the advantage of not only having a simple setup but also the possibility to measure simultaneously five cell parameters on the characterization of TNLCD at high speed.

Lymphatic vessel segmentation in optical coherence tomography by adding U-Net-based CNN for artifact minimization.

The lymphatic system branches throughout the body to transport bodily fluid and plays a key immune-response role. Optical coherence tomography (OCT) is an emerging technique for the noninvasive and label-free imaging of lymphatic capillaries utilizing low scattering features of the lymph fluid. Here, the proposed lymphatic segmentation method combines U-Net-based CNN, a Hessian vesselness filter, and a modified intensity-thresholding to search the nearby pixels based on the binarized Hessian mask. Compared to previous approaches, the method can extract shapes more precisely, and the segmented result contains minimal artifacts, achieves the dice coefficient of 0.83, precision of 0.859, and recall of 0.803.

Quantification of structural and microvascular changes for diagnosing early-stage oral cancer.

Changes in mucosal microvascular networks, called intraepithelial papillary capillary loops (IPCL), are an important key factor for diagnosing early-stage oral cancer in vivo. Nevertheless, there are a lack of tools to quantify these changes objectively. This is the first study to quantify the IPCL changes in vivo to differentiate benign or malignant oral lesions by the optical coherence tomography (OCT) technique. K14-EGFP-miR-211-GFP transgenic mice were inducted by 4-Nitroquinoline-1-oxide to produce oral carcinogenesis in different stages, including normal, premalignancy and cancer. The results showed significant differentiation between benign or malignant lesions by OCT quantitative parameters, including epithelial thickness, IPCL density, radius and tortuosity.

Detection of Oral Dysplastic and Early Cancerous Lesions by Polarization-Sensitive Optical Coherence Tomography.

Detection of oral dysplastic and early-stage cancerous lesions is difficult with the current tools. Half of oral cancers are diagnosed in a late stage. Detection of early stromal change to predict malignant transformation is a new direction in the diagnosis of early-stage oral cancer. The application of new optical tools to image stroma in vivo is under investigation, and polarization-sensitive optical coherence tomography (PS-OCT) is potentially one of those tools. This is a preliminary study to sequentially image oral stromal changes from normal, hyperplasia, and dysplasia to early-stage cancer by PS-OCT in vivo. We used 4-Nitroquinoline-1-oxide drinking water to induce dysplasia and early-stage oral cancer in 19 K14-EGFP-miR-211-GFP transgenic mice. A total of 8 normal, 12 hyperplastic, 11 dysplastic, and 4 early-stage cancerous lesions were enrolled. A new analytic process of PS-OCT imaging was proposed, called an en-face birefringence map. From the birefringence map, the sensitivity, specificity, positive predictive value, and negative predictive values to detect dysplasia and early-stage cancer were 100.00%, 95.00%, 93.75%, and 100.00%, respectively, and the kappa value of these images between two investigators was 0.942. The mean size of malignant lesions detected in this study is 1.66 ± 0.93 mm. This pilot animal study validates the use of PS-OCT to detect small and early-stage oral malignancy with high accuracy and consistency.

Elucidation of microstructural changes in leaves during senescence using spectral domain optical coherence tomography.

Leaf senescence provides a unique window to explore the age-dependent programmed degradation at organ label in plants. Here, spectral domain optical coherence tomography (SD-OCT) has been used to study in vivo senescing leaf microstructural changes in the deciduous plant Acer serrulatum Hayata. Hayata leaves show autumn phenology and change color from green to yellow and finally red. SD-OCT image analysis shows distinctive features among different layers of the leaves; merging of upper epidermis and palisade layers form thicker layers in red leaves compared to green leaves. Moreover, A-scan analysis showed a significant (p < 0.001) decrease in the attenuation coefficient (for wavelength range: 1100-1550 nm) from green to red leaves. In addition, the B-scan analysis also showed significant changes in 14 texture parameters extracted from second-order spatial gray level dependence matrix (SGLDM). Among these parameters, a set of three features (energy, skewness, and sum variance), capable of quantitatively distinguishing difference in the microstructures of three different colored leaves, has been identified. Furthermore, classification based on k-nearest neighbors algorithm (k-NN) was found to yield 98% sensitivity, 99% specificity, and 95.5% accuracy. Following the proposed technique, a portable noninvasive tool for quality control in crop management can be anticipated.

Analog differential-phase detection in optical coherence reflectometer.

A novel differential-phase optical coherence reflectometer (DP-OCR) was proposed using a low-coherence source, integrating it with an analog differential-phase decoding method. In the experiment, the DP-OCR performed a localized surface profile measurement of an optical grating (1200 lp/mm) and demonstrated its ability to measure the translation speed of a tilted mirror. Experimentally, the resolution of the axial displacement of proposed DP-OCR at 185 pm was demonstrated.

Assessment of arterial characteristics in human atherosclerosis by extracting optical properties from polarization-sensitive optical coherence tomography.

In this research, we assessed arterial characteristics in human atherosclerosis by determining both scattering and birefringence properties of vessel tissue from polarization-sensitive optical coherence tomography (PS-OCT) images. We demonstrated that optical characteristics including scattering coefficient (mu(s)), effective anisotropy factor (g(eff)), birefringence coefficient (Delta n), and fast-axis angle (beta) of normal and different kinds of atherosclerotic lesions in human vessels can be quantitatively described. Based on our findings, a quantitative PS-OCT image criterion for plaque characterization was constructed.