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.
Nondestructive three-dimensional (3D) pathology based on high-throughput 3D microscopy holds promise as a complement to traditional hematoxylin and eosin (H&E) stained slide-based two-dimensional (2D) pathology by providing rapid 3D pathological information. However, conventional techniques provided superficial information only due to shallow imaging depth. Herein, we developed open-top two-photon light sheet microscopy (OT-TPLSM) for intraoperative 3D pathology. A two-photon excitation light sheet, generated by 1D scanning of a Bessel beam illuminated the sample and planar imaging was conducted at 400 frames/s max. An imaging depth of 60-100 μm was achieved with long excitation wavelengths, and the image throughput was up to 1 cm2 per 7 min. Cells and extra-cellular matrix were visualized using extrinsic fluorescence and intrinsic second harmonic generation, respectively. OTTPLSM was tested in various human cancer specimens and cancer structures were detected via 3D visualization. OT-TPLSM may have the potential for rapid and precise 3D histopathological examination.
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.
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