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Imaging and quantifying ganglion cells and other transparent neurons in the living human retina

  1. Donald T. Millera
  1. aSchool of Optometry, Indiana University, Bloomington, IN 47405;
  2. bPurdue School of Engineering and Technology, Indiana University–Purdue University Indianapolis, Indianapolis, IN 46202
  1. Edited by David R. Williams, University of Rochester, Rochester, NY, and approved October 18, 2017 (received for review June 30, 2017)

  1. Fig. 2.

    Cellular structures of the inner layers of the retina using AO-OCT. (A) Yellow square at 12–13.5° temporal to the fovea in subject S3 denotes location imaged with AO-OCT. (B) Three-dimensional perspective of registered and averaged AO-OCT volume with green dashed line denoting cross-section of inner retina shown in C. Yellow arrow indicates same GCL soma in C and F. Images shown in DG were extracted at depths of 0, 13, 22, and 46 μm below ILM. Scale bar in G also applies to DF. (D) Surface of ILM. Bright, irregular star-like structures sparsely cover the surface of the ILM and are consistent in appearance with individual astrocyte or microglial cells. (E) A complex web of nerve fiber bundles of varying size disperse across the NFL. Some have a diameter as large as 30 μm (blue arrow), which compares to our previous AO-OCT observations (48). Others are as small as 3 μm, which matches the caliper of a single large GC axon. An arteriole/venule branches on the left. GCL somas appear between the overlying bundles near the image bottom (green arrow). (F) A mosaic of GCL somas of varying size tile the layer. Red arrow points to a large soma. Caliper of arteriole/venule in E is sufficiently large that it extends into the GCL. Note the distinct edges of the vessel walls (blue and white arrows) and the tight abutment of GCL somas. (G) The dense synaptic connections between axons of bipolar cells and dendrites of ganglion and amacrine cells present as a uniform mesh of high spatial frequency irregularities in the IPL. COST, cone outer segment tip; IS/OS, inner segment/outer segment junction; ONL, outer nuclear layer; OPL, outer plexiform layer (Movie S1).

  2. Fig. 3.

    En face images extracted from GCL at increasing retinal eccentricity of subject S4. A mosaic of GCL somas is observed at each eccentricity. (Bottom Right) GC soma density is plotted along the horizontal meridian of the macula. Retinal eccentricity is converted to millimeters to compare with histology data (10). AO-OCT temporal data are the average from four subjects and nasal is from S4. Error bars denote ±1 SD.

  3. Fig. 5.

    Cells at different depths in the same retinal patch of subject S4 as visualized with AO-OCT. (A) Three-dimensional perspective of registered and averaged AO-OCT volume with colored lines denoting retinal depths at which the en face images in BF were extracted. Images depicting individual NF bundles (B), GCL somas (19,162 cells per mm2) (C), suggestive somas of bipolar (green arrow) and displaced GCs (white arrow) near the IPL interface (D), cone photoreceptors (16,341 cells per mm2) (E), and RPE cells (4,893 cells per mm2) (F). Black arrows in CF indicate the same blood vessel and its shadow. The en face images were extracted from volumes acquired at 3–4.5° retinal eccentricity with system focus shifted axially to maximize sharpness of the cell layer of interest.

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