June 2017
Volume 17, Issue 7
Open Access
OSA Fall Vision Meeting Abstract  |   June 2017
Imaging invisible cells: new advances in adaptive optics reveal structure of the translucent retinal cells of the inner retina
Author Affiliations
  • Andres Guevara-Torres
    University of Rochester
Journal of Vision June 2017, Vol.17, 2. doi:10.1167/17.7.2
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      Jesse Schallek, Aby Joseph, Vigneshwar Subramanian, Andres Guevara-Torres; Imaging invisible cells: new advances in adaptive optics reveal structure of the translucent retinal cells of the inner retina. Journal of Vision 2017;17(7):2. doi: 10.1167/17.7.2.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

While the advent of adaptive optics ophthalmoscopy has provided microscopic resolution in the living eye, the majority of neurons in the mammalian retina have evaded detection. This is because most retinal cells are highly translucent, allowing photons to travel through the neural retina with minimal scatter and absorption before reaching the photoreceptors. While retinal translucency is beneficial for vision, it poses a challenge for imaging because cells provide weak optical contrast.

Recent advances in non-confocal ophthalmoscopy now reveal cells that were once hidden by their translucency. By comparing directional cell scatter, we provide detailed images of: ganglion cells, horizontal cells, multi-laminar photoreceptor somata, red blood cells, platelets, white blood cells and putative sub-cellular organelles in the living eye without contrast agents.

We optimized the non-confocal contrast by integrating “split-detection” capabilities (Scoles et al. 2014) into an adaptive optics scanning light ophthalmoscope (AOSLO) custom built for the mouse eye (Guevara-Torres et al. 2015). The split-detection approach compares directional light scatter in the retina. The imaged point spread function of the AOSLO was bisected by a knife-edge prism and relayed into two, phase-locked photomultiplier tubes. The left and right half of the imaged retinal point spread function was digitally subtracted to remove the mutual light information common in the two channels. C57BL/6J mice were anesthetized and imaged with and without contrast agents applied to validate cell type.

Imaged ganglion cells were confirmed by imaging tagged Thy-1 fluorescent cells. Leukocytes were confirmed with fluorescent labeling of nuclei with acridine orange. Red blood cells, platelets, photoreceptor somata and horizontal cells matched the known morphology, density and/or topography in the mouse retina. This new catalog of cells has greatly expanded the number of cell types that may be studied in the living mammalian retina in health and disease.

Meeting abstract presented at the 2016 OSA Fall Vision Meeting

Scoles, Drew, Sulai, Yusufu N., Langlo, Christopher S., Fishman, Gerald A., Curcio, Christine A., Carroll, Joseph, Dubra, Alfredo 2014. “In Vivo Imaging of Human Cone Photoreceptor Inner Segments.” Investigative Ophthalmology & Visual Science 55(7): 4244– 51 doi:10.1167/iovs.14-14542 [CrossRef] [PubMed]
Guevara-Torres, A., Williams, D. R., Schallek, J. B. 2015a. “Imaging Translucent Cell Bodies in the Living Mouse Retina without Contrast Agents.” Biomedical Optics Express 6(6): 2106. doi:10.1364/BOE.6.002106
Footnotes
 Supported by the Research to Prevent Blindness (RPB) Career Development Award, an Unrestricted Grant to the Flaum Eye Institute from RPB, New York, NIH Kirschstein NRSA Postdoctoral Fellowship F32EY023496 and the RPB Stein Innovation Award.
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