April 2010
Volume 51, Issue 13
ARVO Annual Meeting Abstract  |   April 2010
In vivo Two-Photon Imaging of Macaque Retina
Author Affiliations & Notes
  • J. J. Hunter
    Center for Visual Science,
    University of Rochester, Rochester, New York
  • B. Masella
    Institute of Optics,
    University of Rochester, Rochester, New York
  • A. Dubra
    Flaum Eye Institute,
    University of Rochester, Rochester, New York
  • R. Sharma
    Institute of Optics,
    University of Rochester, Rochester, New York
  • G. Palczewska
    Polgenix, Inc., Cleveland, Ohio
  • K. Palczewski
    Department of Pharmacology, Case Western Reserve University, Cleveland, Ohio
  • D. R. Williams
    Center for Visual Science,
    University of Rochester, Rochester, New York
  • Footnotes
    Commercial Relationships  J.J. Hunter, Polgenix, Inc., F; B. Masella, Polgenix, Inc., F; A. Dubra, Polgenix, Inc., F; U.S. Patent #20080007693, P; R. Sharma, Polgenix, Inc., F; G. Palczewska, Polgenix, Inc., E; K. Palczewski, Polgenix, Inc., E; D.R. Williams, Polgenix, Inc., F; Optos, C; U.S. Patents #5,777,719, #5,949,521, #6,095,651, #6,379,005, #6,948,818, #6,199,986, #6,299,311, #6,827,444, #6,264,328, #6,338,559, #20080007693, P.
  • Footnotes
    Support  Polgenix, Inc.; NIH Grants EY001319, EY004367, EY014375, EY007125, EY09339; STC NSF AST-9876783; Research to Prevent Blindness; Burroughs Welcome Fund; Klaus Tschira Fdn.; European Life Scientist Org.
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 3451. doi:https://doi.org/
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      J. J. Hunter, B. Masella, A. Dubra, R. Sharma, G. Palczewska, K. Palczewski, D. R. Williams; In vivo Two-Photon Imaging of Macaque Retina. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3451. doi: https://doi.org/.

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

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Purpose: : In vivo

Methods: : Images were acquired with an adaptive optics scanning laser ophthalmoscope equipped with a dispersion-compensated Ti:Sapph laser source tuned to 730 nm. Adaptive optics produced a near-diffraction limited excitation spot on the retina, reducing the incident flux required to generate a two-photon signal. The emission between 390 nm and 550 nm was collected without descanning from a 1.5º square retinal region. Eye motion during imaging was computed from simultaneously acquired high signal-to-noise ratio (SNR) near-infrared reflectance images, allowing registration of up to 18,000 low SNR two-photon images captured over 12 minutes.

Results: : We obtained a fluorescence signal from the photoreceptor/RPE complex in the living primate eye that was proportional to the square of the incident power, confirming the two-photon nature of the fluorescence. The light levels required for two-photon imaging produced no retinal damage observable with AO imaging, fundus photography and fluorescein angiography. Two-photon fluorescence images appear as a regular array of spots whose size and separation increases with increasing retinal eccentricity in a manner consistent with the cone mosaic. However, the light appears to originate from a layer below the photoreceptors, probably the RPE, since the two-photon signal increases following photopigment bleaching.

Conclusions: : In vivo

Keywords: imaging/image analysis: non-clinical • photoreceptors • retinal pigment epithelium 

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