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In Vivo Vascular Injury Readouts in Mouse Retina to Promote Reproducibility

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Visualizing the invisible: inner plexiform layer stratification with conventional spectral-domain optical coherence

Ricardo Luz Leitão Guerra^1,2, Luiz Roisman^2,3, Jay S Duker^4,5

¹Retina Department, Leitão Guerra - Oftalmologia, Salvador, Brazil.

International Journal of Retina and Vitreous

|June 15, 2025

Summary

This study demonstrates that spectral-domain OCT (SD-OCT) with optimized imaging and grayscale inversion can visualize the five sub-bands of the inner plexiform layer (IPL), crucial for visual processing. This accessible method enhances retinal circuit analysis, previously requiring specialized equipment.

Keywords:

Inner plexiform layer Optical coherence tomography Retinal imaging Spectral-domain

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Published on: August 4, 2018

Area of Science:

Ophthalmology
Neuroscience
Medical Imaging

Background:

The inner plexiform layer (IPL) is vital for retinal visual processing, comprising five sub-bands (S1-S5) that separate ON and OFF pathways.
Previously, visualizing these IPL sub-layers required experimental high-resolution OCT (HR-OCT) or visible-light OCT (VIS-OCT), limiting clinical application.
This study introduces a novel method for visualizing IPL stratification using standard spectral-domain OCT (SD-OCT) technology.

Purpose of the Study:

To demonstrate that IPL stratification can be visualized using commercially available SD-OCT devices.
To evaluate the effectiveness of optimized imaging parameters and grayscale inversion for IPL sub-layer visualization.
To compare the performance of SD-OCT with swept-source OCT (SS-OCT) for visualizing IPL strata.

Main Methods:

Retrospective analysis of macular OCT images from three healthy individuals.
Imaging performed using SD-OCT devices (Nidek RS3000 Advance, Zeiss Cirrus 6000) and one SS-OCT device (Topcon Triton DRI).
High-density B-scans with noise reduction were analyzed in standard and inverted grayscale modes, with varying scan sizes (12mm, 6mm, 3mm).

Main Results:

Inverted grayscale imaging successfully revealed five distinct IPL sub-bands in all SD-OCT cases, particularly in the parafoveal region.
Hyperreflective dots, likely the superficial capillary plexus, were observed near IPL-1.
A 3-mm scan protocol offered superior sub-layer differentiation compared to 12-mm scans, while SS-OCT images did not resolve the five IPL strata.

Conclusions:

IPL stratification is identifiable with conventional SD-OCT through optimized imaging and grayscale inversion, challenging previous assumptions.
This technique enhances retinal circuit analysis using standard clinical technology, offering new diagnostic capabilities.
SD-OCT shows promise for ophthalmology research and monitoring neurodegenerative diseases, whereas SS-OCT is less suitable for this specific application.