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New wrinkles in retinal densitometry.

Benjamin D Masella¹, Jennifer J Hunter², David R Williams³

¹The Institute of Optics, University of Rochester, Rochester, New York, United States Center for Visual Science, University of Rochester, Rochester, New York, United States.

Investigative Ophthalmology & Visual Science

|October 16, 2014

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Retinal densitometry measures retinal function, but interpretation is complicated by reflectance changes unrelated to photopigment. A new spatial normalization method using adaptive optics scanning light ophthalmoscopy minimizes these effects for more accurate optical density measurements.

Area of Science:

Ophthalmology
Retinal Physiology
Optical Imaging

Background:

Retinal densitometry offers objective insights into retinal function.
Interpretation challenges arise from retinal reflectance changes not solely due to photopigment depletion.
Understanding these confounding factors is crucial for accurate retinal function assessment.

Purpose of the Study:

To investigate factors influencing retinal reflectance during densitometry.
To develop and evaluate a method to minimize the impact of non-photopigment-related reflectance changes.
To improve the accuracy of optical density measurements in the retina.

Main Methods:

Utilized an adaptive optics scanning light ophthalmoscope (AOSLO) for photoreceptor reflectance measurements in monkeys.

Keywords:

adaptive optics dark adaptation densitometry intrinsic signals rhodopsin regeneration

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Measured reflectance changes after photopigment bleaching using 514-nm light.

Compared multiple normalization techniques, including a novel spatial normalization method, to extract photopigment optical density.

Main Results:

Identified stimulus-related reflectance fluctuations at 794 nm in both rods and cones, independent of photopigment absorptance.
These fluctuations were significant enough to preclude normalization using infrared reflectance.
The spatial normalization method and confocal AOSLO yielded unexpectedly low apparent rhodopsin density values.

Conclusions:

Confirms that photopigment absorption changes are not the sole source of retinal reflectance variation.
Suggests stray light predominantly originates from retinal layers near the photoreceptors, impacting densitometry.
Despite challenges, the refined method remains a valuable tool for objective retinal function measurement.