Three-Dimensional Analysis of Peeled Internal Limiting Membrane Using Focused Ion Beam/Scanning Electron Microscopy

Area of Science:

• Ophthalmology
• Cell Biology
• Surgical Science

Background:

• Internal limiting membrane (ILM) peeling is a common surgical step in vitrectomy.
• Müller cells are crucial for retinal structure and function.
• Assessing ILM peeling's impact on Müller cells is vital for surgical safety.

Purpose of the Study:

• To reevaluate the effect of ILM peeling during vitrectomy on Müller cell damage.
• To analyze the ultrastructure of the ILM using focused ion beam/scanning electron microscopy (FIB/SEM).

Main Methods:

• Examined 12 ILMs from macular hole and idiopathic epiretinal membrane surgeries using FIB/SEM.
• Analyzed the three-dimensional structures of the ILM.
• Quantified cell fragments on the ILM surface.

Main Results:

• Epiretinal membrane (ERM) group showed significantly higher numbers (12.85/100 μm²) and total volume (10.45 μm³/100 μm²) of cell fragments compared to the macular hole (MH) group (5.07/100 μm² and 3.54 μm³/100 μm²).
• P-values were 0.0024 for cell number and 0.0022 for total volume, indicating statistical significance.
• No significant difference in mean cell fragment volume between MH and ERM groups.

Conclusions:

• All examined ILMs had cell fragments on their retinal surface.
• ERM peeling resulted in more cell fragments than MH peeling, suggesting increased inner retinal damage risk.
• FIB/SEM is an effective tool for three-dimensional quantitative analysis of the ILM.