Efficient Assessment of Tumor Vascular Shutdown by Photodynamic Therapy on Orthotopic Pancreatic Cancer Using High-Speed Wide-Field Waterproof Galvanometer Scanner Photoacoustic Microscopy.

To identify the vascular alteration by photodynamic therapy (PDT), the utilization of high-resolution, high-speed, and wide-field photoacoustic microscopy (PAM) has gained enormous interest. The rapid changes in vasculature during PDT treatment and monitoring of tumor tissue activation in the orthotopic pancreatic cancer model have received limited attention in previous studies. Here, a fully two-axes waterproof galvanometer scanner-based photoacoustic microscopy (WGS-PAM) system was developed for in vivo monitoring of dynamic variations in micro blood vessels due to PDT in an orthotopic pancreatic cancer mouse model. The photosensitizer (PS), Chlorin e6 (Ce6), was utilized to activate antitumor reactions in response to the irradiation of a 660 nm light source. Microvasculatures of angiogenesis tissue were visualized on a 40 mm2 area using the WGS-PAM system at 30 min intervals for 3 h after the PDT treatment. The decline in vascular intensity was observed at 24.5% along with a 32.4% reduction of the vascular density at 3 h post-PDT by the analysis of PAM images. The anti-vascularization effect was also identified with fluorescent imaging. Moreover, Ce6-PDT increased apoptotic and necrotic markers while decreasing vascular endothelial growth factor (VEGF) expression in MIA PaCa-2 and BxPC-3 pancreatic cancer cell lines. The approach of the WGS-PAM system shows the potential to investigate PDT effects on the mechanism of angiographic dynamics with high-resolution wide-field imaging modalities.

A depth-resolved quantitative evaluation method for non-carious cervical lesions treatment with optical coherence tomography.

OBJECTIVES: The aim of this study is prognostic assessment of surface smoothness and the presence of internal bubbles after treatment of non-cancerous cervical lesions (NCCLs) using optical coherence tomography (OCT). METHODS: After treatment with NCCLs, cross-sectional images of the lesion parts of the sample were non-invasively acquired and analyzed. The surface smoothness between tooth and resin, resin and cemento-enamel junction, and the presence bubble inside resin was confirmed. In addition, using an algorithm that distinguishes between resin and dental structure based on OCT cross-sectional images, we quantitatively analyzed the amount of resin used in treating NCCLs and acquired 3D images. RESULTS: The inner structure of the resin in each sample was checked, and the presence of bubbles was confirmed. In addition, the resin sections were separated from the tomographic images acquired by OCT to visualize 3D images. The volume of resin used in the treatment part of each NCCLs samples was quantitatively analyzed as 3.7216 ∼ 14.889 mm3. CONCLUSIONS: OCT is able to measure not only the surface abrasion provided by existing intraoral scanner, but also the size and depth location of interal bubbles, which is distinctive advantage of our method. Based on our results, OCT is a significant tool for qualitative and quantitative analysis of dental NCCLs treatment before and after treatment. CLINICAL SIGNIFICANCE: The study used OCT, a non-destructive diagnostic, to reveal the structure of the resin and the location and size of bubbles after NCCLs treatment. These findings could be golden standard in determining the prognosis of NCCLs treatment.

A concept to detect a subgingival finish line using an intraoral optical coherence tomography system: A clinical report.

This clinical report introduces an approach for detecting the supragingival finish line by penetrating the teeth and gingival tissue using optical coherence tomography (OCT) technology. This approach was used in 3 patients who underwent tooth preparation with a subgingival finish line. Consequently, the subgingival finish line, typically challenging to discern clearly in intraoral scans, was identifiable in the OCT image.

Skin pore imaging using spectral-domain optical coherence tomography: a case report.

Sebum is an important component of the skin that has attracted attention in many fields, including dermatology and cosmetics. Pore expansion due to sebum on the skin can lead to various problems. Therefore, it is necessary to analyze the morphological characteristics of sebum. In this study, we used optical coherence tomography (OCT) to evaluate facial sebum areas. We obtained the OCT maximum amplitude projection (MAP) image and a cross-sectional image of skin pores in the facial area. Subsequently, we detected the sebum in skin pores using the detection algorithm of the ImageJ software to quantitatively determine the size of randomly selected pores in the proposed MAP images. Additionally, the pore size was analyzed by acquiring images before and after facial sebum extraction. According to our research, facial sebum can be morphologically described using the OCT system. Since OCT imaging enables specific analysis of skin parameters, including pores and sebum, skin analysis employing OCT could be an effective method for further research.

Barium sulfate and pigment admixture for photoacoustic and x-ray contrast imaging of the gut.

Significance: X-ray imaging is frequently used for gastrointestinal imaging. Photoacoustic imaging (PAI) of the gastrointestinal tract is an emerging approach that has been demonstrated for preclinical imaging of small animals. A contrast agent active in both modalities could be useful for imaging applications. Aim: We aimed to develop a dual-modality contrast agent comprising an admixture of barium sulfate with pigments that absorb light in the second near-infrared region (NIR-II), for preclinical imaging with both x-ray and PAI modalities. Approach: Eleven different NIR-II dyes were evaluated after admixture with a 40% w/v barium sulfate mixture. The resulting NIR-II absorption in the soluble fraction and in the total mixture was characterized. Proof-of-principle imaging studies in mice were carried out. Results: Pigments that produced more uniform suspensions were assessed further for photoacoustic contrast signal at a wavelength of 1064 nm that corresponds to the output of the Nd:YAG laser used. Phantom imaging studies demonstrated that the pigment-barium sulfate mixture generated imaging contrast in both x-ray and PAI modalities. The optimal pigment selected for further study was a cyanine tetrafluoroborate salt. Ex-vivo and whole-body mouse imaging demonstrated that photoacoustic and x-ray contrast signals co-localized in the intestines for both imaging modalities. Conclusion: These data demonstrate that commercially-available NIR-II pigments can simply be admixed with barium sulfate to generate a dual-modality contrast agent appropriate for small animal gastrointestinal imaging.

Development of a deviated focusing-based optical coherence microscope with a variable depth of focus for high-resolution imaging.

The aim of this study was to develop an optically deviated focusing-based variable depth-of-focus (DOF) oriented optical coherence microscopy (OCM) system to improve the DOF in high-resolution and precise focused imaging. In this study, an approach of varying beam diameter using deviated focusing was employed in the sample arm to enhance the DOF and to confirm precise focusing in OCM imaging. The optically deviated focusing technique was used to vary the focal point and DOF by altering the sample arm beam. The efficacy of the variable DOF imaging approach utilizing an optimized sample arm was confirmed by tissue-level imaging, where OCM images with varying DOF were obtained using deviated focusing. Experimentally confirmed lateral resolution of 2.19 µm was sufficient for the precise non-invasive visualization of abnormalities of fruit specimens. Thus, the proposed variable DOF-OCM system can be an alternative for precisely focused, high-resolution, and variable DOF imaging by improving the DOF in minimum lateral resolution variation.

Dental diagnosis for inlay restoration using an intraoral optical coherence tomography system: A case report.

PATIENTS: The patient was a 32-year-old man who underwent amalgam restoration of the mandibular right second molar. An amalgam restoration fracture was diagnosed by intraoral optical coherence tomography (OCT), and pulp exposure was examined during cavity preparation. Subsequently, a definitive ceramic restoration was fabricated, and the marginal fit in the oral cavity was evaluated using the OCT system. DISCUSSION: The existing OCT system cannot acquire images inside the oral cavity because of the large probe size. However, the proposed intraoral OCT system can access the prostheses in the mandibular right second molar. Therefore, dental diagnosis for restoration treatment with dental prosthesis fracture, marginal gap, and pulp exposure after tooth preparation is possible using the proposed intraoral OCT system. CONCLUSIONS: The use of the intraoral OCT system improved dental diagnosis by allowing the dentist to confirm quantitative values through cross-sectional images, rather than that by determining a treatment plan after visual dental diagnosis.

Three-dimensional reconstructing undersampled photoacoustic microscopy images using deep learning.

Spatial sampling density and data size are important determinants of the imaging speed of photoacoustic microscopy (PAM). Therefore, undersampling methods that reduce the number of scanning points are typically adopted to enhance the imaging speed of PAM by increasing the scanning step size. Since undersampling methods sacrifice spatial sampling density, by considering the number of data points, data size, and the characteristics of PAM that provides three-dimensional (3D) volume data, in this study, we newly reported deep learning-based fully reconstructing the undersampled 3D PAM data. The results of quantitative analyses demonstrate that the proposed method exhibits robustness and outperforms interpolation-based reconstruction methods at various undersampling ratios, enhancing the PAM system performance with 80-times faster-imaging speed and 800-times lower data size. The proposed method is demonstrated to be the closest model that can be used under experimental conditions, effectively shortening the imaging time with significantly reduced data size for processing.

Optical Coherence Tomography as a Non-Invasive Tool for Plant Material Characterization in Agriculture: A Review.

Characterizing plant material is crucial in terms of early disease detection, pest control, physiological assessments, and growth monitoring, which are essential parameters to increase production in agriculture and prevent unnecessary economic losses. The conventional methods employed to assess the aforementioned parameters have several limitations, such as invasive inspection, complexity, high time consumption, and costly features. In recent years, optical coherence tomography (OCT), which is an ultra-high resolution, non-invasive, and real-time unique image-based approach has been widely utilized as a significant and potential tool for assessing plant materials in numerous aspects. The obtained OCT cross-sections and volumetrics, as well as the amplitude signals of plant materials, have the capability to reveal vital information in both axial and lateral directions owing to the high resolution of the imaging system. This review discusses recent technological trends and advanced applications of OCT, which have been potentially adapted for numerous agricultural applications, such as non-invasive disease screening, optical signals-based growth speed detection, the structural analysis of plant materials, and microbiological discoveries. Therefore, this review offers a comprehensive exploration of recent advanced OCT technological approaches for agricultural applications, which provides insights into their potential to incorporate OCT technology into numerous industries.

On-field optical imaging data for the pre-identification and estimation of leaf deformities.

Visually nonidentifiable pathological symptoms at an early stage are a major limitation in agricultural plantations. Thickness reduction in palisade parenchyma (PP) and spongy parenchyma (SP) layers is one of the most common symptoms that occur at the early stage of leaf diseases, particularly in apple and persimmon. To visualize variations in PP and SP thickness, we used optical coherence tomography (OCT)-based imaging and analyzed the acquired datasets to determine the threshold parameters for pre-identifying and estimating persimmon and apple leaf abnormalities using an intensity-based depth profiling algorithm. The algorithm identified morphological differences between healthy, apparently-healthy, and infected leaves by applying a threshold in depth profiling to classify them. The qualitative and quantitative results revealed changes and abnormalities in leaf morphology in addition to disease incubation in both apple and persimmon leaves. These can be used to examine how initial symptoms are influenced by disease growth. Thus, these datasets confirm the significance of OCT in identifying disease symptoms nondestructively and providing a benchmark dataset to the agriculture community for future reference.

Target ischemic stroke model creation method using photoacoustic microscopy with simultaneous vessel monitoring and dynamic photothrombosis induction.

The ischemic stroke animal model evaluates the efficacy of reperfusion and neuroprotective strategies for ischemic injuries. Various conventional methods have been reported to induce the ischemic models; however, controlling specific neurological deficits, mortality rates, and the extent of the infarction is difficult as the size of the affected region is not precisely controlled. In this paper, we report a single laser-based localized target ischemic stroke model development method by simultaneous vessel monitoring and photothrombosis induction using photoacoustic microscopy (PAM), which has minimized the infarct size at precise location with high reproducibility. The proposed method has significantly reduced the infarcted region by illuminating the precise localization. The reproducibility and validity of suggested method have been demonstrated through repeated experiments and histological analyses. These results demonstrate that our method can provide the ischemic stroke model closest to the clinical pathology for brain ischemia research from inducement, occurrence mechanisms to the recovery process.

Non-destructive morphological observation of anatomical growth process in Haemaphysalis Longicornis tick specimens using optical coherence tomography.

BACKGROUND: Ticks are known as the representatives of hematophagous arachnids. They cause various tick-borne diseases, such as severe fever with thrombocytopenia syndrome (SFTS) and Lyme disease. To understand the mechanism of virus infection caused by ticks, morphology for the anatomical characteristics of crucial organs has been widely studied in acarological fields. The conventional methods used for tick observation have inevitable limitations. Dissection is the standard method to obtain the morphological information, and complex microscopy methods were utilized alternatively. OBJECTIVE: The study goal is to obtain the morphological information of ticks in different growth stages non-invasively. METHODS: Optical coherence tomography (OCT) is employed to acquire structural images of various internal organs without damage for observing the growth process of larva, nymph, and adult in Haemaphysalis longicornis ticks in real-time. RESULTS: Various internal organs, such as salivary glands, rectal sac, genital aperture, and anus, were well-visualized by the OCT enface and cross-sectional images, and the variation in size of these organs in each growth stage was compared quantitatively. CONCLUSIONS: Based on the obtained results, we confirmed the potential feasibility of OCT as a non-destructive real-time tool for morphological studies in acarology. Further research using OCT for acarological applications can include monitoring the growth process of ticks in terms of structural changes and investigating morphological differences between normal and virus-infected tick specimens.

Non-Invasive Optical Coherence Tomography Data-Based Quantitative Algorithm for the Assessment of Residual Adhesive on Bracket-Removed Dental Surface.

The aim of this study was to quantitatively assess the residual adhesive on orthodontic ceramic bracket-removed dental surface. In orthodontic process, ceramic bracket was repeated debonding physically, then the adhesive remained on the dental surface. The residual adhesive caused a lack of adhesive strength between dental and ceramic bracket. Since commonly used adhesive in orthodontics is translucent, residual adhesive is hard to be detected with conventional microscopes. Therefore, 1310 nm center wavelength swept-source OCT system based on laboratory customized image processing algorithm was used for the precise detection of residual adhesive on tooth surface. The algorithm separates residual adhesive from dental surface by comparing the height of adjacent B-scan images, while providing color-scaled images emphasizing the thickness information of residual adhesive. Finally, the acquired results were compared with microscopic and adhesive remnant index scoring gold standards, while the comparison confirmed the potential merits and the improvements of the proposed method over gold standards.

Micron-scale human enamel layer characterization after orthodontic bracket debonding by intensity-based layer segmentation in optical coherence tomography images.

In clinical orthodontic practice, fixed brackets are widely used for tooth movement and adjustments. Although years of research and development have improved the workability of fixed orthodontic brackets, there are still controversies regarding its plausible destructive influence on the enamel surface of tooth. This, in turn, makes the quantitative assessment of the enamel surface after specific orthodontic treatment procedures important in order to opt for the most effective treatment procedure. Through this study, we show the practical applicability of optical coherence tomography (OCT) as a non-ionizing and nondestructive assessment tool for measuring enamel loss after each step of orthodontic bracket bonding. Two-dimensional and volumetric OCT images are used for the evaluation of the tooth enamel. From the depth intensity profile analysis of cross-sectional OCT images, the changes in the individual internal layer thickness are calculated. A software algorithm was developed to evaluate the structural connectivity in the enamel for analyzing enamel loss on the tooth surface and for detecting enamel abrasion. An intensity-based layer segmentation algorithm is also developed to analyze and evaluate enamel wear in the tooth after each step. Using the proposed algorithms, the total enamel present after each treatment procedure was measured and tabulated for analysis.

Optical Coherence Tomography for Quantifying Human Cutaneous Chronic Graft-versus-Host Disease.

Chronic graft-versus-host disease (cGVHD) is the most common cause of nonrelapse mortality after allogeneic hematopoietic cell transplantation (alloHCT). Cutaneous cGVHD is characterized by thickening of the skin and connective tissues, causing discomfort and limited mobility. Current assessment of these skin lesions is based on physical examination of their thickening, pinchability, and movability. Optical coherence tomography (OCT) is a noninvasive, high-resolution technique using near-infrared light to interrogate tissues and image the microstructure without the use of contrast agents. We determined the applicability of OCT to human cutaneous cGVHD. Seven patients with varying degrees of cutaneous cGVHD, including 3 controls who underwent autologous HCT were prospectively examined using the cGVHD Skin (Vienna) Scale and imaged with OCT. Analysis of OCT images and clinical exams revealed that stratum corneum thickness, epidermal thickness, and depth of light transmission were correlated with cutaneous cGVHD severity in the hands, forearms, upper arms, legs, thighs, and upper back (P ≤ .03). Longitudinal OCT changes during cGVHD treatment paralleled clinical changes in the arm and upper back. OCT changes were observed in the absence of clinical changes. OCT imaging reflects the severity of cutaneous cGVHD and can be used to follow these lesions. OCT may facilitate the design of therapeutic trials in cGVHD by providing a quantitative measurement of cGVHD severity. Additional studies are needed.

Integrated Quad-Scanner Strategy-Based Optical Coherence Tomography for the Whole-Directional Volumetric Imaging of a Sample.

Whole-directional scanning methodology is required to observe distinctive features of an entire physical structure with a three dimensional (3D) visualization. However, the implementation of whole-directional scanning is challenging for conventional optical coherence tomography (OCT), which scans a limited portion of the sample by utilizing unidirectional and bidirectional scanning methods. Therefore, in this paper an integrated quad-scanner (QS) strategy-based OCT method was implemented to obtain the whole-directional volumetry of a sample by employing four scanning arms installed around the sample. The simultaneous and sequential image acquisition capabilities are the conceptual key points of the proposed QS-OCT method, and were implemented using four precisely aligned scanning arms and applied in a complementary way according to the experimental criteria. To assess the feasibility of obtaining whole-directional morphological structures, a roll of Scotch tape, an ex vivo mouse heart, and kidney specimens were imaged and independently obtained tissue images at different directions were delicately merged to compose the 3D volume data set. The results revealed the potential merits of QS-OCT-based whole-directional imaging, which can be a favorable inspection method for various discoveries that require the dynamic coordinates of the whole physical structure.

A preliminary study of post-progressive nail-art effects on in vivo nail plate using optical coherence tomography-based intensity profiling assessment.

Nail beautification is a widely applied gender independent practice. Excessive nail beautifications and nail-arts have a direct impact on the nail structure and can cause nail disorders. Therefore, the assessment of post-progressive nail-art effects on the nail is essential to maintain optimal nail health and to avoid any undesirable disorders. In this study, in vivo nails were examined in control stage, with a nail-art stage, and after removing the nail-art stage using a 1310 nm spectral-domain optical coherence tomography (SD-OCT) system. The acquired cross-sectional OCT images were analyzed by a laboratory customized signal processing algorithm to obtain scattered intensity profiling assessments that could reveal the effects of nail beautification on the nail plate. The formation and progression of cracks on the nail plate surface were detected as an effect of nail beautification after 72 h of nail-art removal. Changes in backscattered light intensity and nail plate thickness of control and art-removed nails were quantitatively compared. The results revealed the potential feasibility of the developed OCT-based inspection procedure to diagnose post-progressive nail-art effects on in vivo nail plate, which can be helpful to prevent nail plate damages during art removal through real-time monitoring of the boundary between the nail plate and nail-art. Besides nail-art effects, the developed method can also be used for the investigation of nail plate abnormalities by examining the inconsistency of internal and external nail plate structure, which can be diagnosed with both qualitative and quantitative assessments from a clinical perspective.

Multi-directional Morphological Assessment of Single Bacterial Colonies Through Non-invasive Optical Imaging.

A bacterium in bacterial colony is a basal component of bacterial studies and is therefore of considerable importance. The morphological characteristics of a single colony have been widely used as the standard to identify diverse bacterial species. However, the conventional methods for obtaining morphological information, such as microscopic techniques, cannot provide tomographic views. This study utilizes spectral-domain optical coherence tomography (SD-OCT) to observe both external and internal structures for single colonies. OCT imaging provided a clear top (projection) and lateral (cross-sectional) images, which can identify distinct features in the fifteen different bacterial species examined. Through the acquired OCT images, the elevation type in each genus was confirmed, and the quantitative measurements for morphology including a diameter, height, and elevation angle were achieved. Moreover, the OCT raw data was used to build three-dimensional images to observe outer appearances in all direction. As a result, the averaged angles of 7.92°, 14.40°, 6.78°, and 11.92° of the genus Hymenobacter, Spirosoma, Bacillus, and Deinococcus, respectively, were calculated. In conclusion, we have demonstrated that OCT is a highly feasible, non-invasive method for the identification of bacterial species, providing detailed morphological and visual information. It thus has strong potential for applications in bacteriology, as well as biomedical and food sciences.

Ultra-stiff compressed collagen for corneal perforation patch graft realized by in situ photochemical crosslinking.

Despite the potential of a collagen construct, consisting of a major extracellular matrix component of the native cornea, as a patch graft to treat the corneal perforation, there has still been difficulty in acquiring sufficient mechanical properties for clinical availability. This study developed a novel in situ photochemical crosslinking (IPC)-assisted collagen compression process, namely, the IPC-C2 process, to significantly enhance the mechanical properties of the collagen construct for the development of a collagenous patch graft. For the first time, we found that compressed collagen construct was rapidly rehydrated in an aqueous solution, which inhibited effective riboflavin-mediated photochemical crosslinking for mechanical improvement. The IPC-C2 process was designed to concurrently induce the physical compaction and photochemical crosslinking of a compressed collagen construct, thereby avoiding the loosening of collagen fibrillar structure during rehydration and ultimately improving crosslinking efficiency. Hence, the suggested IPC-C2 process could fabricate a collagen construct with a high collagen density (∼120-280 mg ml-1) and ∼103-fold increased mechanical properties (an elastic modulus of up to ∼29 MPa and ultimate tensile strength of ∼8 MPa) compared with collagen gel. This construct can then be used as a clinically applicable collagenous patch graft. With sufficient mechanical strength for surgical suture and the controllable thickness for patient specificity, the potential of the fabricated IPC-compressed collagen construct for clinical applications was demonstrated by using an in vivo rabbit corneal perforation model. It effectively protected aqueous humor leakage and maintained the integrity of the eye globe without an additional complication.

Optical Interferometric Fringe Pattern-Incorporated Spectrum Calibration Technique for Enhanced Sensitivity of Spectral Domain Optical Coherence Tomography.

Depth-visualizing sensitivity can be degraded due to imperfect optical alignment and non-equidistant distribution of optical signals in the pixel array, which requires a measurement of the re-sampling process. To enhance this depth-visualizing sensitivity, reference and sample arm-channeled spectra corresponding to different depths using mirrors were obtained to calibrate the spectrum sampling prior to Fourier transformation. During the process, eight interferogram patterns corresponding to point spread function (PSF) signals at eight optical path length differences were acquired. To calibrate the spectrum, generated intensity points of the original interferogram were re-indexed towards a maximum intensity range, and these interferogram re-indexing points were employed to generate a new lookup table. The entire software-based process consists of eight consecutive steps. Experimental results revealed that the proposed method can achieve images with a high depth-visualizing sensitivity. Furthermore, the results validate the proposed method as a rapidly performable spectral calibration technique, and the real-time images acquired using our technique confirm the simplicity and applicability of the method to existing optical coherence tomography (OCT) systems. The sensitivity roll-off prior to the spectral calibration was measured as 28 dB and it was halved after the calibration process.