Viet Nhat Hung Le1, Yanhong Hou1, Jens Horstmann1,2, Felix Bock1,3, Claus Cursiefen1,3. 1. Department of Ophthalmology, University Hospital of Cologne, Cologne, Germany. 2. Cluster of Excellence: Cellular Stress Responses in Aging-associated Diseases, CECAD, University of Cologne, Cologne, Germany. 3. Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany.
Abstract
PURPOSE: Corneal lymphatic vessels are clinically invisible because of their thin walls and clear lymph fluid. There is no easy and established method for in vivo imaging of corneal lymphatic vessels so far. In this study, we present a novel approach to visualize corneal lymphatic vessels in vivo by injecting intrastromal fluorescein sodium. METHODS: Six- to eight-week-old female BALB/c mice were used in the mouse model of suture-induced corneal neovascularization. Two weeks after the suture placement, fluorescein sodium was injected intrastromally. The fluorescein, taken up by the presumed lymphatic vessels, was then tracked using a clinically used Spectralis HRA + OCT device. Immunohistochemistry staining with specific lymphatic marker LYVE-1 and pan-endothelial marker CD31 was used to confirm the indirect lymphangiography findings. RESULTS: By injecting fluorescein intrastromally, both corneal blood and lymphatic vessels were detected. While the lymphatic vessels were visible as bright vessel-like structures using HRA, the blood vessels appeared as dark networks. Fluorescein-labeled lymphatic vessels were colocalized with LYVE-1 in immunohistochemically stained sections of the same specimen. CONCLUSIONS: Corneal lymphatic vessels can be easily imaged in vivo in the murine model using intrastromal fluorescein injection.
PURPOSE:Corneal lymphatic vessels are clinically invisible because of their thin walls and clear lymph fluid. There is no easy and established method for in vivo imaging of corneal lymphatic vessels so far. In this study, we present a novel approach to visualize corneal lymphatic vessels in vivo by injecting intrastromal fluorescein sodium. METHODS: Six- to eight-week-old female BALB/c mice were used in the mouse model of suture-induced corneal neovascularization. Two weeks after the suture placement, fluorescein sodium was injected intrastromally. The fluorescein, taken up by the presumed lymphatic vessels, was then tracked using a clinically used Spectralis HRA + OCT device. Immunohistochemistry staining with specific lymphatic marker LYVE-1 and pan-endothelial marker CD31 was used to confirm the indirect lymphangiography findings. RESULTS: By injecting fluorescein intrastromally, both corneal blood and lymphatic vessels were detected. While the lymphatic vessels were visible as bright vessel-like structures using HRA, the blood vessels appeared as dark networks. Fluorescein-labeled lymphatic vessels were colocalized with LYVE-1 in immunohistochemically stained sections of the same specimen. CONCLUSIONS:Corneal lymphatic vessels can be easily imaged in vivo in the murine model using intrastromal fluorescein injection.
Authors: Yifan Wu; Young Jin Seong; Kin Li; Dongwon Choi; Eunkyung Park; George H Daghlian; Eunson Jung; Khoa Bui; Luping Zhao; Shrimika Madhavan; Saren Daghlian; Patill Daghlian; Desmond Chin; Il-Taeg Cho; Alex K Wong; Martin Heur; Sandy Zhang-Nunes; James C Tan; Masatsugu Ema; Tina T Wong; Alex S Huang; Young-Kwon Hong Journal: JCI Insight Date: 2020-07-09
Authors: Uta Gehlsen; Daniela Stary; Martina Maass; Katarina Riesner; Gwen Musial; Michael E Stern; Olaf Penack; Philipp Steven Journal: Int J Mol Sci Date: 2021-06-08 Impact factor: 5.923