Junsoo Lee

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.

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.

Mice and rats are rodent specimens commonly used in multidisciplinary research. Specifically, vasculature imaging of rodents has been widely performed in preclinical studies using various techniques, such as computed tomography, magnetic resonance imaging, and ultrasound imaging. Photoacoustic CT (PACT) is a noninvasive, nonionizing optical imaging technique derived from photoacoustic tomography and benefits from using intrinsic endogenous contrast agents to produce three-dimensional volumetric data from images. In this study, a commercial PACT device was employed to assess the cervicothoracic vasculature of mouse and rat specimens, which has rarely been examined using PACT, under two conditions with depilation and skin incision. Various blood vessels, including the common carotid artery, internal/external jugular veins, cranial vena cava, internal thoracic vein, and mammary, were identified in the acquired PACT images. The difference between the depilated and skin-incised specimens also revealed the presence of branches from certain blood vessels and specific anatomical features such as the manubrium of the sternum. This study presents detailed PACT images observing the cervicothoracic vasculature of rodent specimens and is expected to be used as a reference for various preclinical experiments on mice and rats.

Photoacoustic imaging (PAI) is a hybrid non-invasive imaging technique used to merge high optical contrast and high acoustic resolution in deep tissue. PAI has been extensively developed by utilizing its advantages that include deep imaging depth, high resolution, and label-free imaging. As a representative implementation of PAI, photoacoustic microscopy (PAM) has been used in preclinical and clinical studies for its micron-scale spatial resolution capability with high optical absorption contrast. Several handheld and portable PAM systems have been developed that improve its applicability to several fields, making it versatile. In this study, we developed a laboratory-customized, two-axis, waterproof, galvanometer scanner-based handheld PAM (WP-GVS-HH-PAM), which provides an extended field of view (14.5 × 9 mm²) for wide-range imaging. The fully waterproof handheld probe enables free movement for imaging regardless of sample shape, and volume rate and scanning region are adjustable per experimental conditions. Results of WP-GVS-HH-PAM-based phantom and in vivo imaging of mouse tissues (ear, iris, and brain) confirm the feasibility and applicability of our system as an imaging modality for various biomedical applications.

The accurate screening of otitis media (OM) lies in clarifying the numerous confounding and quantitative factors that are discovered during primary inspections. Increased awareness about bacterial biofilms and inflammation has allowed researchers to develop a better understanding of the bacterial infections that occur in the middle ear. In this study, four live guinea pigs were inoculated with Streptococcus pneumonia to induce OM-related inflammatory changes. Since optical techniques have been effectively used for diagnosis in medicine, low-coherence interferometry-based optical coherence tomography (OCT) was employed for depth-resolved high-resolution data screening. Multiple locations of the tympanic membrane (TM), mastoid mucosa, and round window membrane were examined to assess inflammatory changes. We performed qualitative assessments and thickness quantifications and investigated the variations due to adverse inflammatory affects. The findings of the present study provide a better understanding of S. pneumonia bacteria caused inflammatory changes in middle ear and it further provides a fundamental platform for future clinical utility.

Optical fiber is widely used in optical coherence tomography (OCT) to propagate light precisely with low attenuation and low dispersion. However, the total optical path length within the optical fiber varies in accordance with changes of the temperature. This leads changes in the total optical travel path of the interfering signals and results in shifting of OCT image position to an unintended depth pixel value. In this paper, we presented the temperature-based automatic path length compensating method in OCT to limit the external temperature effect and control the image position in micro-scale without manual movement of optical components. By utilizing developed hardware and software of automatic temperature control system, the external temperature of optical fiber is precisely regulated that evokes thermal expansion and finally changes the physical length of fiber, which is main mechanism of temperature-based path length compensating method. The effectiveness of the presented method was verified by two-dimensional OCT images of mirror and in vivo retina. The obtained results confirmed the path length variance due to temperature change is computable and can be regulated in real-time for whole pixel range of OCT image. Therefore, the proposed temperature-based path length compensating method can be used as an alternative method to precisely control the position of OCT image, while eliminating the effect of external temperature and apply to effectively configuring compact optical systems.

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.

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.

On-field in situ inspection of Marssonina coronaria infected apple blotch for apple leaf specimens was demonstrated using compact backpack-type optical coherence tomography (OCT). Simultaneously, the acquired OCT results were precisely matched with a widely used agricultural plant pathogen inspection technique called loop-mediated isothermal amplification (LAMP) to confirm on-field applicability of the compact backpack-type OCT. Specimens were examined for 8 weeks. Automated Amplitude-scan (A-scan) depth profiling and post-processed infected tissue boundary detection from acquired OCT images were performed to investigate the increase in the internal layer gaps of the leaf specimens resulting from the disease. Clearly identifiable morphological difference between healthy and infected-suspected specimens was observed through the OCT images, which were well correlated with LAMP results. OCT-LAMP correlations and confirmed feasibility study results conclude that the compact backpack-type OCT diagnosing modality can be effective and extensively applicable for various novel agricultural discoveries.