Supplemental Figure 1: High-resolution AO images of the cone mosaics in Figure 5 in normal
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2 Supplemental Figure 1: High-resolution AO images of the cone mosaics in Figure 5 in normal control subjects (NCS), carriers of choroideremia (CAR) and choroideremia patients (CHM) showing the locations of each cone determined by direct counting and used for measurements of cone density. Cones were identified by author JM through a semi-automated custom Matlab 26, 27 program. Cone densities are shown for each image, and all densities measured fell within the previously reported normal range. Manual cone identification still leaves room for error through misidentifying or missed cones. This underscores the need for well-designed studies to test the repeatability, reliability, and variability of cone analysis with multiple observers and for more accurate, automated cone identification programs to alleviate the need for manual intervention. 2
3 Supplemental Figure 2: High-resolution AO images of the cone mosaic at approximately 0.5 mm along each meridian in choroideremia patients (CHM). Cone densities for each of the images in the two left columns fell within the previously reported normal range (Supplemental Figure 3). 32 Cone densities for each of the images in the two right columns fell below the previously reported normal range. 32 Scale bar 20 um. 3
4 Supplemental Figure 3: AO images of the cone mosaics in Supplemental Figure 2 at 0.5mm in choroideremia patients (CHM) showing the locations of each cone determined by direct counting and used for measurements of cone density. 4
5 Supplemental Figure 4: IR and OCT images from the left eye of symptomatic carrier showing large regions of RPE atrophy in the macula, including the foveal region. The atrophic border exhibits the classic scalloped edges characteristic of choroideremia. OCT shows loss of outer retinal layers in regions of atrophy, thinned RPE and indistinguishable interdigitation layer 5
6 in regions of relatively intact retina, abrupt borders at the edge of atrophy, interlaminar bridges, and outer retinal tubulation. Supplemental video 1: High resolution AOSLO images recorded at various focus levels in patient where bubble-like features are found adjacent to the centrally intact retina. This is the same location as shown in Figure 10 patient By adjusting the focus of the AO system, the bubble-like features came into best focus approximately 0.4D posterior to the plane of best focus for the cone photoreceptors. This image series was obtained using a custom built AOSLO as previously described. 40 The OCT for in Figure 10 shows that the bubble-like features co-locate with the hypo-reflective space in the en face translational directions; this through focus video this gives further evidence that the bubble-like features also co-locate with the hypo-reflective space in the choroid in depth. Supplemental Table 1: Normal Control Patient Data * Number of retinal locations at which cone density was measured (Number of locations where cone density was within/higher/lower than the normal range reported by Song et al. 32 ) Supplemental Table 2: Choroideremia Carrier Patient Data * Eye was excluded from study due to AOSLO imaging artifact Number of retinal locations at which cone density was measured (Number of locations where cone density was within/higher/lower than the normal range reported by Song et al. 32 ) Carrier was symptomatic 6
7 Supplementary Table 1: Normal Control Patient Data Patient ID Age Axial Length (mm) Cone Density Measured * OD OS OD OS (5/0/1) 12 (12/0/0) (7/1/1) 8 (8/0/0) (11/1/0) 4 (4/0/0) (6/0/0) 5 (4/1/0) (7/0/0) 6 (6/0/0) (1/0/0) 4 (4/0/0) (5/0/0) 9 (9/0/0) (6/0/1) 8 (8/0/0) * Number of retinal locations at which cone density was measured (Number of locations where cone density was within/higher/lower than the normal range reported by Song et al. )
8 Supplementary Table 2: Choroideremia Carrier Patient Data Patient ID Age Axial Length (mm) Visual Acuity Cone Density Measured OD OS OD OS OD OS /20 20/20 5 (4/0/1) 1 (1/0/0) (10/1/0) 11 (11/0/0) /20 20/20 1 (1/0/0) 4 (4/0/0) * * 4 (4/0/0) /20 20/20 4 (4/0/0) 3 (2/0/1) /20 20/20 6 (5/0/1) 4 (4/0/0) (10/1/0) 11 (11/0/0) /20 20/20 2 (2/0/0) /20 20/20 10 (9/1/0) 2 (2/0/0) /20 20/20 11 (11/0/0) 9 (8/0/1) (10/1/0) 9 (8/1/0) /20 20/20 3 (3/0/0) 7 (6/1/0) /20 20/20 * 3 (2/0/1) dB 7.67dB 5 (3/1/1) 5 (5/0/0) (4/0/0) 1 (1/0/0) /20 20/20 2 (2/0/0) 3 (3/0/0) (1/1/0) 1 (1/0/0) /20 20/20 2 (2/0/0) * * Eye was excluded from study due to AOSLO imaging artifact Number of re nal loca ons at which cone density was measured (Number of locations where cone density was within/higher/lower than the normal range reported by Song et al. ) Carrier was symptoma c Data not available Visual acuity data not available but Humphrey Visual field data was available. Central 30 2 Threshold Test, Mean Deviation db
Micropulse Duty Cycle. # of eyes (20 ms) Total spots (200 ms)
Micropulse Duty Cycle Total spots (2 ms) # of eyes (2 ms) Total spots (2 ms) % 269 44 3 47% 9 4 4 25% 3 5 4 4 5% 2 4 3 5 2% 5 2 NA NA 9% 2 4 6% NA NA 57 2 5% 4 5 6 3 3% 39 5 35 5 # of eyes (2 ms) Supplemental
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