Adaptive optics approach measures the basis of color vision

Adaptive optics scanning laser ophthalmoscopy enabled studying color vision by probing photoreceptors in the human eye.

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Using a technique called adaptive optics scanning laser ophthalmoscopy, researchers at University Eye Hospital Bonn and colleagues studied color vision by probing photoreceptors (individual sensory cells) in the human eye. Their results confirm that the photoreceptor cells of the retina are especially sensitive to colors corresponding to their visual pigments, even when stimulated in isolation. Sensitivity of tested photoreceptors varied, depending on which cell classes were located in their immediate neighborhood.

Related: Adaptive optics open a new frontier of in vivo subcellular imaging

When the light is switched on in a dark room, color vision sets in. "This not only makes the world more colorful," says Dr. Wolf M. Harmening, who heads an Emmy Noether research group at Bonn University Eye Hospital and is one of the lead authors of a paper describing the work, "color also allows spatial detail to become apparent that has proven vital for survival over the course of evolution."

Some predator camouflage can only be identified through color. Poisonous animals and plants also provide warning signals through color. That human color vision emerges from three independent channels within the retina is well established in vision science literature. By stimulating individual photoreceptor cells in living subjects, Harmening and co-lead author William S. Tuten from the University of California, Berkeley, together with colleagues from U.S. universities in Seattle, WA, and Birmingham, AL, have now shown on a cellular scale how the human retina conveys color signals.

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Dr. Wolf M. Harmening (left) from University Eye Hospital Bonn and Dr. William S. Tuten (right) from the University of California, Berkeley. (Photo copyright: Rolf Müller/Ukom-UKB)

To do this, the researchers used adaptive optics scanning laser ophthalmoscopy, which can examine and stimulate the human retina noninvasively. The novel method employs a combination of a laser and a very high-resolution microscope that can map individual sensory cells in the retina. The research team has now used this ophthalmoscope to study vision in the retinas of two human subjects.

According to common theory, all color stimuli can be formed by mixing the primary colors red, green, and blue. While rod photoreceptors are specialized for seeing in the dark, cone photoreceptors convey color vision. They carry light sensitive pigments specialized to absorb wavelengths near the primary colors, the basis of trichromatic vision.

The researchers initially mapped the cone mosaic on the subjects' retinas by measuring light absorption for certain wavelengths in each photoreceptor. In this way, they were able to determine the sensory cells' identity, or class, within the framework of trichromacy. By reducing the intensity of the stimulation light, the researchers were then able to determine a detection threshold in each cone, at which light was just barely seen by the subjects. "This is important because we could use the sensitivity of each cell to determine how overall perception is governed by the contribution of individual cones," Harmening explains.

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Researchers at the University of Bonn and the University of California, Berkeley analyzed the sensitivity of photoreceptor cells. (Copyright: William S. Tuten/Wolf M. Harmening)

The sensitivity of single cells also depended on the immediate neighboring cells. "If a cone sensitive to red light is surrounded by cells that are more sensitive to green, this cone is more likely to behave like a green cone," Harmening says. Studying visual processing of color is complex in part because the brain does not receive raw data from individual photoreceptors, but rather an already preprocessed retinal signal. "Spatial and color information of individual cones is modulated in the complex network of the retina, with lateral information spreading through what are known as horizontal cells," Harmening explains.

Their finding supports previous assumptions about color vision. "What's new is that we can now study vision on the most elementary level, cell by cell," Harmening says. Conventional tests of vision use stimuli that necessarily activate hundreds to thousands photoreceptor cells at the same time. Harmening emphasizes that cellular-scale retinal computation such as the proximity effect has important implications, for basic and clinical research. "When the basis of vision is understood better, we open avenues for new diagnoses and treatments in case of retinal disease," Harmening says. The novel single cell approach offers access to new findings in ophthalmology.

Full details of the work appear in The Journal of Neuroscience.

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