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.

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.

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.

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.

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.

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.

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.

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.

Fully waterproof two-axis galvanometer scanner for enhanced wide-field optical-resolution photoacoustic microscopy.

A large field-of-view and fast scanning of photoacoustic microscopy (PAM) relatively have been difficult to obtain due to the water-drowned structure of the system for the transmission of ultrasonic signals. Researchers have widely studied the achievement of a waterproof scanner for dynamic biological applications with a high-resolution and high signal-to-noise ratio. This Letter reports a novel, to the best of our knowledge, waterproof galvanometer scanner-based PAM system with a successfully attainable ${9.0}\;{\rm mm} \times {14.5}\;{\rm mm}$9.0mm×14.5mm scan region, amplitude scan rate of 40 kHz, and spatial resolution of 4.9 µm. The in vivo characterization of a mouse brain in intact-skull microvascular visualization demonstrated its capability in biomedical imaging and is anticipated to be an effective technique for various preclinical and clinical studies.

In Situ Characterization of Micro-Vibration in Natural Latex Membrane Resembling Tympanic Membrane Functionally Using Optical Doppler Tomography.

Non-invasive characterization of micro-vibrations in the tympanic membrane (TM) excited by external sound waves is considered as a promising and essential diagnosis in modern otolaryngology. To verify the possibility of measuring and discriminating the vibrating pattern of TM, here we describe a micro-vibration measurement method of latex membrane resembling the TM. The measurements are obtained with an externally generated audio stimuli of 2.0, 2.2, 2.8, 3.1 and 3.2 kHz, and their respective vibrations based tomographic, volumetric and quantitative evaluations were acquired using optical Doppler tomography (ODT). The micro oscillations and structural changes which occurred due to diverse frequencies are measured with sufficient accuracy using a highly sensitive ODT system implied phase subtraction method. The obtained results demonstrated the capability of measuring and analyzing the complex varying micro-vibration of the membrane according to implied sound frequency.

Biocompatibility evaluation of bioprinted decellularized collagen sheet implanted in vivo cornea using swept-source optical coherence tomography.

Corneal transplantation by full-thickness penetrating keratoplasty with human donor tissue is a widely accepted treatment for damaged or diseased corneas. Although corneal transplantation has a high success rate, a shortage of high-quality donor tissue is a considerable limitation. Therefore, bioengineered corneas could be an effective solution for this limitation, and a decellularized extracellular matrix comprises a promising scaffold for their fabrication. In this study, three-dimensional bioprinted decellularized collagen sheets were implanted into the stromal layer of the cornea of five rabbits. We performed in vivo noninvasive monitoring of the rabbit corneas using swept-source optical coherence tomography (OCT) after implanting the collagen sheets. Anterior segment OCT images and averaged amplitude-scans were acquired biweekly to monitor corneal thickness after implantation for 1 month. The averaged cornea thickness in the control images was 430.3 ± 5.9 µm, while the averaged thickness after corneal implantation was 598.5 ± 11.8 µm and 564.5 ± 12.5 µm at 2 and 4 weeks, respectively. The corneal thickness reduction of 34 µm confirmed the biocompatibility through the image analysis of the depth-intensity profile base. Moreover, hematoxylin and eosin staining supported the biocompatibility evaluation of the bioprinted decellularized collagen sheet implantation. Hence, the developed bioprinted decellularized collagen sheets could become an alternative solution to human corneal donor tissue, and the proposed image analysis procedure could be beneficial to confirm the success of the surgery.

Fast Industrial Inspection of Optical Thin Film Using Optical Coherence Tomography.

An application of spectral domain optical coherence tomography (SD-OCT) was demonstrated for a fast industrial inspection of an optical thin film panel. An optical thin film sample similar to a liquid crystal display (LCD) panel was examined. Two identical SD-OCT systems were utilized for parallel scanning of a complete sample in half time. Dual OCT inspection heads were utilized for transverse (fast) scanning, while a stable linear motorized translational stage was used for lateral (slow) scanning. The cross-sectional and volumetric images of an optical thin film sample were acquired to detect the defects in glass and other layers that are difficult to observe using visual inspection methods. The rapid inspection enabled by this setup led to the early detection of product defects on the manufacturing line, resulting in a significant improvement in the quality assurance of industrial products.

High-speed and high-SNR photoacoustic microscopy based on a galvanometer mirror in non-conducting liquid.

Optical-resolution photoacoustic microscopy (OR-PAM), a promising microscopic imaging technique with high ultrasound resolution and superior optical sensitivity, can provide anatomical, functional, and molecular information at scales ranging from the microvasculature to single red blood cells. In particular, real-time OR-PAM imaging with a high signal-to-noise ratio (SNR) is a prerequisite for widespread use in preclinical and clinical applications. Although several technical approaches have been pursued to simultaneously improve the imaging speed and SNR of OR-PAM, they are bulky, complex, not sensitive, and/or not actually real-time. In this paper, we demonstrate a simple and novel OR-PAM technique which is based on a typical galvanometer immersed in non-conducting liquid. Using an opto-ultrasound combiner, this OR-PAM system achieves a high SNR and fast imaging speed. It takes only 2 seconds to acquire a volumetric image with a wide field of view (FOV) of 4 × 8 mm2 along the X and Y axes, respectively. The measured lateral and axial resolutions are 6.0 and 37.7 µm, respectively. Finally, as a demonstration of the system's capability, we successfully imaged the microvasculature in a mouse ear in vivo. Our new method will contribute substantially to the popularization and commercialization of OR-PAM in various preclinical and clinical applications.