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.
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.
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