Related Experiment Video
Updated: Jun 4, 2025

Whole Mount Immunofluorescent Staining of the Neonatal Mouse Retina to Investigate Angiogenesis In vivo
Published on: July 9, 2013
Yin-Yang Staining Technique to Create a Nonstained Internal Limiting Membrane Flap to Cover Large Idiopathic Macular
Yao Ni1, Shuwen Xing1, Baizhou Chen1
1State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
Abstract:
Purpose: To describe a modified technique for negative and positive (Yin-Yang) staining of the internal limiting membrane (ILM) to create a nonstained ILM flap that covers large idiopathic macular holes (MHs). Methods: Consecutive patients with large idiopathic MHs (>400 μm) were prospectively included in the study. After the central vitreous was removed, a droplet of triamcinolone acetonide was injected, covering the MH and surrounding area. Subsequently, indocyanine green (ICG) was injected to stain the outer area of the ILM, followed by creation of a flap from the temporal stained area. Finally, the nonstained ILM flap was inverted to cover the MH. The main outcomes included the best-corrected visual acuity (BCVA), macular contour, and integrity of the outer retina. Results: This study comprised 31 patients (31 eyes). Of the eyes, 28 (90.3%) achieved primary MH closure. The mean minimum linear diameter and base diameter of the MH were 593 ± 119 μm and 1082 ± 242 μm, respectively. At the 6-month follow-up, 12 eyes (38.7%) and 9 eyes (29.0%) had regained a U-shaped or V-shaped macular contour, respectively. In addition, the mean logMAR BCVA improved from 1.06 ± 0.30 preoperatively to 0.56 ± 0.31 (P < .001). Twenty-one eyes (67.7%) and 16 eyes (51.6%) had regained integrity of the external limiting membrane and ellipsoid zone, respectively, at the 6-month follow-up. Conclusions: The modified ILM staining technique using triamcinolone acetonide and ICG sequentially is a safe and effective method of creating a nonstained ILM flap that covers large MHs and prevents the foveal area from coming into direct contact with ICG.
More Related Videos
10:14Author Spotlight: Ex Vivo OCT-Based Multimodal Imaging of Human Donor Eyes for Research into Age-Related Macular Degeneration
Published on: May 26, 2023
12:01Techniques for Processing Eyes Implanted With a Retinal Prosthesis for Localized Histopathological Analysis
Published on: August 2, 2013