Intratissue activity imaging of an alveolar organoid with three-dimensional dynamic optical coherence tomography
February 1, 2023 | Microscopy, Other Applications | Damietta University, Medical University of Vienna, University of TsukubaAntonia Lichtenegger, Ibrahim Abd El-Sadek, Kiriko Tomita, Pradipta Mukherjee, Rion Morishita, Shuichi Makita, Tetsuharu Nagamoto, Toshio Suzuki, Yiheng Lim, Yoshiaki Yasuno, Yuki Yamamoto
An organoid is a three-dimensional (3D) in vitro cell culture emulating human organs. We applied 3D dynamic optical coherence tomography (DOCT) to visualize the intratissue and intracellular activities of human induced pluripotent stem cells (hiPSCs)-derived alveolar organoids in normal and fibrosis models. 3D DOCT data were acquired with a 840-nm spectral domain optical coherence tomography with axial and lateral resolutions of 3.8 {\mu}m (in tissue) and 4.9 {\mu}m, respectively. The DOCT images were obtained by the logarithmic-intensity-variance (LIV) method, which is sensitive to the signal fluctuation magnitude. The LIV images revealed cystic structures surrounded by high-LIV borders and mesh-like structures with low LIV. The former may be alveoli with a highly dynamics epithelium, while the latter may be fibroblasts. The LIV images also demonstrated the abnormal repair of the alveolar epithelium.
Optical Coherence Tomography Is a Promising Tool for Zebrafish-Based Research—A Review
January 22, 2023 | Developmental Biology | Medical University of Vienna, University of TsukubaAntonia Lichtenegger, Bernhard Baumann, Yoshiaki Yasuno
The zebrafish is an established vertebrae model in the field of biomedical research. With its small size, rapid maturation time and semi-transparency at early development stages, it has proven to be an important animal model, especially for high-throughput studies. Three-dimensional, high-resolution, non-destructive and label-free imaging techniques are perfectly suited to investigate these animals over various development stages. Optical coherence tomography (OCT) is an interferometric-based optical imaging technique that has revolutionized the diagnostic possibilities in the field of ophthalmology and has proven to be a powerful tool for many microscopic applications. Recently, OCT found its way into state-of-the-art zebrafish-based research. This review article gives an overview and a discussion of the relevant literature and an outlook for this emerging field.
Extending field-of-view of retinal imaging by optical coherence tomography using convolutional Lissajous and slow scan patterns
November 27, 2022 | Ophthalmology | Tokyo Medical & Dental University, University of TsukubaMasahiro Miura, Shinnosuke Azuma, Shuichi Makita, Tatsuo Yamaguchi, Toshihiro Mino, Yoshiaki Yasuno
Optical coherence tomography (OCT) is a high-speed non-invasive cross-sectional imaging technique. Although its imaging speed is high, three-dimensional high-spatial-sampling-density imaging of in vivo tissues with a wide field-of-view (FOV) is challenging. We employed convolved Lissajous and slow circular scanning patterns to extend the FOV of retinal OCT imaging with a 1-µm, 100-kHz-sweep-rate swept-source OCT prototype system. Displacements of sampling points due to eye movements are corrected by post-processing based on a Lissajous scan. Wide FOV three-dimensional retinal imaging with high sampling density and motion correction is achieved. Three-dimensional structures obtained using repeated imaging sessions of a healthy volunteer and two patients showed good agreement. The demonstrated technique will extend the FOV of simple point-scanning OCT, such as commercial ophthalmic OCT devices, without sacrificing sampling density.
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Birefringence-derived scleral artifacts in optical coherence tomography images of eyes with pathologic myopia
November 19, 2022 | Ophthalmology | Tokyo Medical UniversityAtsuya Miki, Hiroyuki Shimizu, Masahiro Miura, Rei Nemoto, Shinnosuke Azuma, Shuichi Makita, Tatsuo Yamaguchi, Toshihiro Mino, Yoshiaki Yasuno
We investigated birefringence-derived scleral artifacts in optical coherence tomography (OCT) images of eyes with pathologic myopia. This study included 76 eyes of 42 patients with pathologic myopia. Five sets of OCT B-scan images of the macula were obtained using commercial swept-source OCT. A dataset of prototype swept-source polarization-diversity OCT images was used to identify polarization-dependent OCT images (i.e., complex averaging of OCT signals from two polarization channels) and polarization-independent OCT images (i.e., intensity averaging of two OCT signals). Polarization-dependent OCT images and commercial OCT images were assessed for the presence of birefringence-derived artifacts by comparison with polarization-independent OCT images. Both polarization-dependent OCT images and commercial OCT images contained scleral vessel artifacts. Scleral vessel artifacts were present in 46 of 76 eyes (60.5%) imaged by polarization-dependent OCT and 17 of 76 eyes (22.4%) imaged by...
Longitudinal investigation of a xenograft tumor zebrafish model using polarization-sensitive optical coherence tomography
September 14, 2022 | Developmental Biology | Harvard University, Massachusetts General Hospital, Medical University of Vienna, University of Tsukuba, University of ViennaAdelheid Wöhrer, Antonia Lichtenegger, Bernhard Baumann, Junya Tamaoki, Lisa Greutter, Lixuan Bian, Makoto Kobayashi, Pradipta Mukherjee, Roxane Licandro, Satoshi Matsusaka, Shuichi Makita, Tomoko Mori, Yoshiaki Yasuno
Breast cancer is a leading cause of death in female patients worldwide. Further research is needed to get a deeper insight into the mechanisms involved in the development of this devastating disease and to find new therapy strategies. The zebrafish is an established animal model, especially in the field of oncology, which has shown to be a promising candidate for pre-clinical research and precision-based medicine. To investigate cancer growth in vivo in zebrafish, one approach is to explore xenograft tumor models. In this article, we present the investigation of a juvenile xenograft zebrafish model using a Jones matrix optical coherence tomography (JM-OCT) prototype. Immunosuppressed wild-type fish at 1-month post-fertilization were injected with human breast cancer cells and control animals with phosphate buffered saline in the tail musculature. In a longitudinal study, the scatter, polarization, and vasculature changes over time were investigated and quantified in control versus t...
Label-free metabolic imaging of non-alcoholic-fatty-liver-disease (NAFLD) liver by volumetric dynamic optical coherence tomography
August 23, 2022 | Uncategorized | University of Tsukuba, University of ViennaAntonia Lichtenegger, Arata Miyazawa, Donny Lukmanto, Ibrahim Abd El-Sadek, Kosuke Okada, Lida Zhu, Pradipta Mukherjee, Rion Morishita, Shinichi Fukuda, Shuichi Makita, Tetsuro Oshika, Toshiharu Yamashita, Yoshiaki Yasuno
Label-free metabolic imaging of non-alcoholic fatty liver disease (NAFLD) mouse liver is demonstrated ex vivo by dynamic optical coherence tomography (OCT). The NAFLD mouse is a methionine choline-deficient (MCD)-diet model, and two mice fed the MCD diet for 1 and 2 weeks are involved in addition to a normal-diet mouse. The dynamic OCT is based on repeating raster scan and logarithmic intensity variance (LIV) analysis that enables volumetric metabolic imaging with a standard-speed (50,000 A-lines/s) OCT system. Metabolic domains associated with lipid droplet accumulation and inflammation are clearly visualized three-dimensionally. Particularly, the normal-diet liver exhibits highly metabolic vessel-like structures of peri-vascular hepatic zones. The 1-week MCD-diet liver shows ring-shaped highly metabolic structures formed with lipid droplets. The 2-week MCD-diet liver exhibits fragmented vessel-like structures associated with inflammation. These results imply that volumetric LIV im...
Non-destructive characterization of adult zebrafish models using Jones matrix optical coherence tomography
March 30, 2022 | Developmental Biology | Medical University of Vienna, University of Tsukuba, University of ViennaAntonia Lichtenegger, Bernhard Baumann, Daisuke Oida, Ibrahim Abd El-Sadek, Kiriko Tomita, Konrad Leskovar, Lida Zhu, Martin Distel, Pradipta Mukherjee, Rion Morishita, Shuichi Makita, Stefanie Kirchberger, Yoshiaki Yasuno
The zebrafish is a valuable vertebrate animal model in pre-clinical cancer research. A Jones matrix optical coherence tomography (JM-OCT) prototype operating at 1310 nm and an intensity-based spectral-domain OCT setup at 840 nm were utilized to investigate adult wildtype and a tumor-developing zebrafish model. Various anatomical features were characterized based on their inherent scattering and polarization signature. A motorized translation stage in combination with the JM-OCT prototype enabled large field-of-view imaging to investigate adult zebrafish in a non-destructive way. The diseased animals exhibited tumor-related abnormalities in the brain and near the eye region. The scatter intensity, the attenuation coefficients and local polarization parameters such as the birefringence and the degree of polarization uniformity were analyzed to quantify differences in tumor versus control regions. The proof-of-concept study in a limited number of animals revealed a significant decrease...
Evaluation of choroidal melanin-containing tissue in healthy Japanese subjects by polarization-sensitive optical coherence tomography
March 15, 2022 | Ophthalmology | Tokyo Medical University, University of TsukubaMasahiro Miura, Shinnosuke Azuma, Shuichi Makita, takuya iwasaki, Tatsuo Yamaguchi, Toshihiro Mino, Yoshiaki Yasuno
In this study, the choroidal melanin content in healthy eyes was evaluated with polarization-sensitive optical coherence tomography (PS-OCT). We evaluated 105 healthy eyes of 105 Japanese subjects. The mean thickness of melanin-containing tissue in the choroid (thickness of MeCh) and the choroidal melanin occupancy rate within a 5-mm circular region from the foveal center were calculated using the degree of polarization uniformity obtained by PS-OCT and compared with the choroidal thickness, patient age, and axial length. To evaluate regional variations, the 5-mm circular region was divided into a center area and an outer ring area, and the outer ring area was further divided into four areas (nasal, temporal, superior, and inferior). The mean thickness of MeCh showed a significant positive correlation with the choroidal thickness. The mean choroidal melanin occupancy rate showed a significant positive correlation with age. The mean choroidal melanin occupancy rate of the center area...
Objective evaluation of choroidal melanin loss in patients with Vogt–Koyanagi–Harada disease using polarization-sensitive optical coherence tomography
March 11, 2022 | Ophthalmology | University of TsukubaHiroshi Goto, Hiroyuki Shimizu, Masahiro Miura, Rei Nemoto, Shinnosuke Azuma, Shuichi Makita, takuya iwasaki, Tatsuo Yamaguchi, Toshihiro Mino, Yoshiaki Yasuno
In this study, sunset glow fundus was evaluated in patients with Vogt-Koyanagi-Harada (VKH) disease using polarization-sensitive optical coherence tomography (PS-OCT). We evaluated 40 VKH eyes (20 patients) and 59 healthy eyes (59 age-matched controls). VKH eyes were divided into three groups according to color fundus images: sunset (17 eyes), potential sunset (13 eyes), and non-sunset (10 eyes). Choroidal melanin thickness (ChMeT) and the choroidal melanin thickness ratio (ChMeTratio) were calculated based on the degree of polarization uniformity from PS-OCT. ChMeT was significantly lower in sunset eyes than in non-sunset or control eyes (P = 0.003). The ChMeTratios of sunset or potential sunset eyes were significantly lower than those of non-sunset or control eyes (P = 0.04). Regional evaluation of ChMeT and the ChMeTratio showed that choroidal depigmentation predominantly occurred in the macula's outer ring area (P = 0.002). The areas under receiver operating characteristic curve...
Multicontrast investigation of in vivo wildtype zebrafish in three development stages using polarization-sensitive optical coherence tomography
January 24, 2022 | Developmental Biology | Medical University of Vienna, University of Tsukuba, University of ViennaAntonia Lichtenegger, Bernhard Baumann, Ibrahim Abd El-Sadek, Junya Tamaoki, Konrad Leskovar, Lida Zhu, Lixuan Bian, Makoto Kobayashi, Pradipta Mukherjee, Yoshiaki Yasuno
Significance: The scattering and polarization characteristics of various organs of in vivo wildtype zebrafish in three development stages were investigated using a non-destructive and label-free approach. The presented results showed a promising first step for the usability of Jones-matrix optical coherence tomography (JM-OCT) in zebrafish-based research. Aim: We aim to visualize and quantify the scatter and polarization signatures of various zebrafish organs for larvae, juvenile, and young adult animals in vivo in a non-invasive and label-free way. Approach: A custom-built polarization-sensitive JM-OCT setup in combination with a motorized translation stage was utilized to investigate live zebrafish. Depth-resolved scattering (intensity and attenuation coefficient) and polarization (birefringence and degree of polarization uniformity) properties were analyzed. OCT angiography (OCT-A) was utilized to investigate the vasculature label-free and non-destructively. Results: The scatter ...
Deep convolutional neural network-based scatterer density and resolution estimators in optical coherence tomography
December 19, 2021 | Uncategorized | University of TsukubaArata Miyazawa, Ibrahim Abd El-Sadek, Kensuke Oikawa, Larina Tzu-Wei Shen, Lida Zhu, Pradipta Mukherjee, Prathan Buranasiri, Satoshi Matsusaka, Shuichi Makita, Thitiya Seesan, Yoshiaki Yasuno
We present deep convolutional neural network (DCNN)-based estimators of the tissue scatterer density (SD), lateral and axial resolutions, signal-to-noise ratio (SNR), and effective number of scatterers (ENS, the number of scatterers within a resolution volume). The estimators analyze the speckle pattern of an optical coherence tomography (OCT) image in estimating these parameters. The DCNN is trained by a large number (1,280,000) of image patches that are fully numerically generated in OCT imaging simulation. Numerical and experimental validations were performed. The numerical validation shows good estimation accuracy as the root mean square errors were 0.23%, 3.65%, 3.58%, 3.79%, and 6.15% for SD, lateral and axial resolutions, SNR, and ENS, respectively. The experimental validation using scattering phantoms (Intralipid emulsion) shows reasonable estimations. Namely, the estimated SDs were proportional to the Intralipid concentrations, and the average estimation errors of lateral a...
Label-free functional and structural imaging of liver microvascular complex in mice by Jones matrix optical coherence tomography
October 10, 2021 | Urology | University of TsukubaArata Miyazawa, Donny Lukmanto, Ibrahim Abd El-Sadek, Kosuke Okada, Lida Zhu, Pradipta Mukherjee, Shinichi Fukuda, Shuichi Makita, Tetsuro Oshika, Toshiharu Yamashita, Yoshiaki Yasuno
We demonstrate label-free imaging of the functional and structural properties of microvascular complex in mice liver. The imaging was performed by a custom-built Jones-matrix based polarization sensitive optical coherence tomography (JM-OCT), which is capable of measuring tissue's attenuation coefficient, birefringence, and tiny tissue dynamics. Two longitudinal studies comprising a healthy liver and an early fibrotic liver model were performed. In the healthy liver, we observed distinctive high dynamics beneath the vessel at the initial time point (0 h) and reappearance of high dynamics at 32-h time point. In the early fibrotic liver model, we observed high dynamics signal that reveals a clear network vascular structure by volume rendering. Longitudinal time-course imaging showed that these high dynamics signals faded and decreased over time.
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Computational multi-directional optical coherence tomography for visualizing the microstructural directionality of the tissue
August 31, 2021 | Uncategorized | University of TsukubaDaisuke Oida, Kensuke Oikawa, Kiriko Tomita, Meng-Tsan Tsai, Shuichi Makita, Tai-Ang Wang, Yoshiaki Yasuno
We demonstrate computational multi-directional optical coherence tomography (OCT) to assess the directional property of tissue microstructure. This method is the combination of phase-sensitive volumetric OCT imaging and post-signal processing. The latter comprises of two steps. The first step is an intensity-directional analysis, which determines the dominant en face fiber orientations. The second step is the phase-directional imaging, which reveals the sub-resolution depth-orientation of the microstructure. The feasibility of the method was tested by assessing muscle and tendon samples. Stripe patterns with several sizes were visualized in the phase-directional images. In order to interpret these images, the muscle and tendon structures were numerically modeled, and the phase-directional images were generated from the numerical model. The similarity of the experimental and numerical results suggested that the stripe patterns correspond to the muscle fiber bundle and its crimping.
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Effect of A-scan rate and interscan interval on optical coherence angiography
March 9, 2021 | Ophthalmology | National Taiwan University, University of TsukubaBo-Huei Huang, Chuan-Bor Chueh, hsiang-chieh lee, Meng-Tsan Tsai, Ting-Hao Chen, Ting-Yen Tsai, Yi-Chun Wu, Yin-Peng Huang, Yoshiaki Yasuno
Optical coherence tomography angiography (OCTA) can provide rapid, volumetric, and noninvasive imaging of tissue microvasculature without the requirement of exogenous contrast agents. To investigate how A-scan rate and interscan time affected the contrast and dynamic range of OCTA, we developed a 1.06-µm swept-source OCT system enabling 100-kHz or 200-kHz OCT using two light sources. After system settings were carefully adjusted, almost the same detection sensitivity was achieved between the 100-kHz and 200-kHz modalities. OCTA of ear skin was performed on five mice. We used the variable interscan time analysis algorithm (VISTA) and the designated scanning protocol with OCTA images reconstructed through the correlation mapping method. With a relatively long interscan time (e.g., 12.5 ms vs. 6.25 ms for 200-kHz OCT), OCTA can identify more intricate microvascular networks. OCTA image sets with the same interscan time (e.g., 12.5 ms) were compared. OCTA images acquired with a 10...
We present a completely label-free three-dimensional (3D) optical coherence tomography (OCT)-based tissue dynamics imaging method for visualization and quantification of the metabolic and necrotic activities of tumor spheroid. Our method is based on a custom 3D scanning protocol that is designed to capture volumetric tissue dynamics tomography images only in a few tens of seconds. The method was applied to the evaluation of a tumor spheroid. The time-course viability alteration and anti-cancer drug response of the spheroid were visualized qualitatively and analyzed quantitatively. The similarity between the OCT-based dynamics images and fluorescence microscope images was also demonstrated.
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