| 녹F-020 |
| Differential interference contrast microscopy 를 이용한 망막신경절세포의 이미징 |
| 서울대학교 의과대학 안과학교실 (1), 한국과학기술연구원(2), 연세대학교 공과대학(3) |
| 김석환(1), 오주영(2), 김유정(1), 김철기(2), 이택진(2), 서민아(2), 이석(2), 우덕하(2), 전성찬(3), 박기호(1), 김재헌(2) |
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목적 : Glaucoma is a progressive optic neuropathy, characterized by the selective loss of retinal ganglion cells (RGCs). Therefore, monitoring the change of number or morphology of RGC is essential for the early detection as well as investigation of pathophysiology of glaucoma. We have previously demonstrated photoreceptor cell imaging with differential interference contrast (DIC) microscopy. In this study, we successfully visualized single RGC using DIC microscopy.
방법 : Since RGC layer is much less reflective than photoreceptor layer, various techniques including the control of light wavelength and bandwidth using a tunable band pass filter were adopted to reduce the chromatic aberration in z-axis for higher and clearer resolution. To verify that the imaged cells were the RGCs, the flat-mounted retina of Sprague-Dawley rat, in which the RGCs were retrogradely labeled with fluorescence, was observed by both fluorescence and DIC microscopies for direct comparison.
결과 : For DIC image, near-infrared light with the wavelength centered at 704 nm and bandwidth of 10 nm was illuminated on the sample to image RGCs. RGCs were clearly shown on DIC image. To verify that the obtained crater-shaped images were RGCs, fluorescence microscopy was also performed. Both DIC and fluorescence images are perfectly matched. Because amacrine cells also exist in RGC layer, some cell images by DIC microscope do not match with cell images by fluorescence microscopy.
결론 : In conclusion, we successfully visualized single RGC with DIC microscopy. Since the DIC is basically label free detection, the conducted experiment shows the possibility for the in-vivo imaging of RGCs.
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