We imaged the retina using the Indiana Adaptive Optics Scanning Laser Ophthalmoscope (AOSLO). Our system uses two deformable mirrors to provide en face, high-resolution images of retinal structures at a 28 Hz frame rate. The wavelength of the sensor light was 850 nm and the imaging wavelength was 820 nm at 50 and 120 W respectively. The confocal pinhole was located in a position conjugate with the retina allowed us to segment one retina plane. Two different confocal apertures of 75 m and 100 m (1.5 and 2 times the Airy disk size) were used to provide different amounts of confocal or scattered light. The imaging area was 1.4 x 1.2 deg which corresponds roughly to 400 x 350 m.
Using the large stroke deformable mirror, which provides the focusing capability of the confocal system, we imaged the same location at different planes. We moved from superficial layers to the retinal pigment epithelium in 0.3 D increments. The range of adjustments included the subjectively best overall image, and focal planes anterior and posterior to this.
We imaged 10 subjects at approximately 7.5 deg temporal from the fovea. A video of individual frames was taken, and the individual frames were dewarped, aligned, and averaged. We measured 10 bright and 10 dim cones for each subject at the 10 depths, with brightness groupings based subjectively on the most superficial location. The function for amount of light reflected differed for the two groups of cones. Reflectivity varied as a function of depth.
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