KEYWORDS: Microscopy, Depth of field, Human subjects, Diseases and disorders, Tolerancing, Real time imaging, Imaging systems, Image deconvolution, Fluorescence microscopy, Eye models
Conjunctival goblet cells (CGCs) are specialized epithelial cells that secrete mucins to form the mucous layer of the protective tear film and by suppressing inflammation. Although CGCs are an important biomarker for diagnosing ocular surface diseases, rapid and noninvasive CGC examination methods have not been available. We have developed a new imaging system, high-speed extended depth-of-field wide-field microscopy with surface tracking, to enable non-contact large-area CGC imaging in human subjects. A novel long-range surface detection method was developed for rapid large-area mosaic imaging with lateral translation. Large-area CGC imaging and density quantification in human subjects was demonstrated. This new imaging system could be useful for noninvasive CGC examination in humans, which could be valuable for precision diagnosis and optimal treatment of ocular surface diseases.
KEYWORDS: Microscopy, Imaging systems, Human subjects, In vivo imaging, Eye, Animal model studies, Visualization, Motion models, Luminescence, Eye models
Mucin secretive conjunctival goblet cells (CGCs) in the eye play important roles in ocular surface homeostasis by forming the mucous layer of the tear film. CGC information is also an important biomarker for diagnosis because CGC loss or dysfunction is observed in various ocular surface diseases. In this study, we developed moxifloxacin-based extended depth-of-field (EDOF) microscopy with surface tracking for non-invasive CGC imaging in awake human subjects. The system had a DOF of 0.8 mm, a field of view (FOV) of 1.3mm x 1.3mm, and imaging speed of 15 fps. The phase detection method was used for real-time surface tracking. Moxifloxacin ophthalmic solution was topically instilled for CGC labeling. Repeated large area imaging of the same conjunctiva in a human subject was demonstrated. MBFM might have the potential for non-invasive CGC examination in patients.
Recently, as the usage of electronic devices increase, modern people suffer from eye diseases. We analyzed goblet cells of wide-field fluorescence microscopy with a deep learning. In this study, we propose to real-time analysis using knowledge distillation using proposed loss function and optimized network. In the proposed method, residual based UNet was used as the teacher network to distill knowledge into lightweight E-Net. We train the student network using pixelwise loss and . The proposed method showed 4% improvements in dice-score compared to the lightweight E-Net, and the processing time was decreased to 68% compared to the case where only the teacher network was performed.
5-ALA-based fluorescence-guided surgery (FGS) is a state-of-the-art treatment for brain tumors, but may miss areas due to its low sensitivity. To improve the sensitivity we developed dual PpIX and Moxifloxacin Fluorescence Confocal Imaging method that allows simultaneous imaging of 5-ALA and moxifloxacin by excitation with a single light source. This method verified by comparing confocal images in glioblastoma specimen. The distribution of cells expressing 5-ALA and moxifloxacin fluorescence signals showed the same correlation. In large-area tumor sample images, PplX showed differences in signal intensity, but moxifloxacin was constant. These results show the potential to improve sensitivity than with 5-ALA alone.
KEYWORDS: Microscopy, Luminescence, Motion models, Imaging systems, Image resolution, Real time imaging, Performance modeling, In vivo imaging, Image filtering, Human subjects
Mucin secretive conjunctival goblet cells (CGCs) in the eye are important for tear film stability and ocular surface health. Because CGC dysfunction is associated with various ocular surface diseases, non-invasive CGC examination will be of great help in the diagnosis and treatment. In this study, we developed a high-speed moxifloxacin-based extended depth-of-field microscopy for real-time CGC examination. The performance was demonstrated by high-speed CGC imaging of both mouse and rabbit models, in vivo. The imaging was insensitive to breathing motion, and the image resolution was sufficient to resolve individual CGCs in rabbit models.
Mucin secretive conjunctival goblet cells (CGCs) play important roles for ocular surface homeostasis by forming the mucous layer of tear film, so CGC examination is important for diagnosis of various ocular surface diseases. Here we show that CGCs can be non-invasively imaged in real time and in high contrasts in animal models by moxifloxacin-based fluorescence microscopy (MBFM) using moxifloxacin antibiotic ophthalmic solution for specific CGC labeling. Newly developed MBFM was applied to both disease mouse and rabbit models and it detected CGC damage and recovery via longitudinal imaging. These results showed that MBFM has potentials for non-invasive CGC examination.
PAL-XFEL has operated general user program successfully from 2017 in both hard (2.5 – 14 keV) and soft (250 – 1300 eV) X-ray beamlines. Particularly in Soft X-ray Scattering and Spectroscopy (SSS) beamline, X-ray absorption and emission spectroscopy (XAS / XES) endstation has conducted general user programs.
In SSS beamline, second experimental instrument, Resonant Soft X-ray Scattering (RSXS) endstation, was built and open from 2020. This endstation mainly supports optical laser pump (800-nm-wavelength and harmonics) soft X-ray probe (400-1300 eV, covers 3d transition metal L2,3-edge, O K-edge, and part of 5f rare-earth element M4,5-edge) time-resolved RSXS experiments, with a time resolution better than 100 fs. Within less than a year, there have been several successful experiments, mainly in 3d transition metal compounds which are important magnetic and strongly correlated materials. The details of the endstation and selected experimental results will be introduced in the presentation.
Surgical resection is the primary treatment for malignant brain tumors. This procedure has a dilemma—aggressive surgical resection tends to extend patient survival; however, it also increases the risk of neurological deficiencies. Current medical imaging methods are not sensitive and their interpretation largely depend on surgeon’s impression. High-speed cellular imaging method by using clinically applicable moxifloxacin was demonstrated for fast and sensitive tumor-detection. The detailed cytoarchitecture of brain tumor mouse model and malignant human brain tumors was revealed. This study showed the potential and feasibility of moxifloxacin-based confocal microscopy as a surgery-guiding method for tumor removal.
Dermal filler injections are common skin cosmetic procedure, but intravascular injection of fillers can lead to various complications. Therefore, real-time imaging of vessels and fillers will be helpful to lower the complications. In this study, angiographic optical coherence tomography (A-OCT) was used for real-time visualization of vessels and fillers. This method was demonstrated in an in vivo mouse model by monitoring filler injection in the tail. The locations of main artery and fillers were identified by A-OCT imaging before and after injection. Our study suggests that the real-time angiographic OCT can serve as a useful tool for filler injection guide.
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