Imaging of single retinal ganglion cell with differential interference contrast microscopy

Title
Imaging of single retinal ganglion cell with differential interference contrast microscopy
Authors
우덕하이석김재헌이택진김철기서민아Juyeong OhYu Jeong KimSeong Chan JunKi-Ho ParkSeok Hwan Kim
Keywords
Ganglion cell imaging; retina; differential interference contrast; label free
Issue Date
2017-01
Publisher
SPIE Photonics West 2017
Citation
VOL 10045, NO 21-90
Abstract
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. Since RGC layer is transparent and hyporeflective, the direct optical visualization of RGCs has not been successful so far. Therefore, glaucoma evaluation mostly depends on indirect diagnostic methods such as the evaluation of optic disc morphology or retinal nerve fiber layer thickness measurement by optical coherence tomography. We have previously demonstrated single photoreceptor cell imaging with differential interference contrast (DIC) microscopy. Herein, we successfully visualized single RGC using DIC microscopy. 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. We have confirmed that the cell images obtained by fluorescence microscopy were perfectly matched with cell images by DIC microscopy. As conclusion, we have visualized single RGC with DIC microscopy, and confirmed with fluorescence microscopy
URI
https://pubs.kist.re.kr/handle/201004/65678
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KIST Publication > Conference Paper
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