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Lu RW, Li YC, Ye T, et al. Two-photon excited autofluorescence imaging of freshly isolated frog retinas. Biomed Opt Express. 2011;2(6):1494-503doi: 10.1364/BOE.2.001494.
Lu, R. W., Li, Y. C., Ye, T., Strang, C., Keyser, K., Curcio, C. A., & Yao, X. C. (2011). Two-photon excited autofluorescence imaging of freshly isolated frog retinas. Biomedical optics express, 2(6), 1494-503. https://doi.org/10.1364/BOE.2.001494
Lu, Rong-Wen, et al. "Two-photon excited autofluorescence imaging of freshly isolated frog retinas." Biomedical optics express vol. 2,6 (2011): 1494-503. doi: https://doi.org/10.1364/BOE.2.001494
Lu RW, Li YC, Ye T, Strang C, Keyser K, Curcio CA, Yao XC. Two-photon excited autofluorescence imaging of freshly isolated frog retinas. Biomed Opt Express. 2011 Jun 01;2(6):1494-503. doi: 10.1364/BOE.2.001494. Epub 2011 May 11. PMID: 21698013; PMCID: PMC3114218.
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Biomed Opt Express. 2011 Jun 01;2(6):1494-503. doi: 10.1364/BOE.2.001494. Epub 2011 May 11.

Two-photon excited autofluorescence imaging of freshly isolated frog retinas.

Biomedical optics express

Rong-Wen Lu, Yi-Chao Li, Tong Ye, Christianne Strang, Kent Keyser, Christine A Curcio, Xin-Cheng Yao

PMID: 21698013 PMCID: PMC3114218 DOI: 10.1364/BOE.2.001494

Abstract

The purpose of this study was to investigate cellular sources of autofluorescence signals in freshly isolated frog (Rana pipiens) retinas. Equipped with an ultrafast laser, a laser scanning two-photon excitation fluorescence microscope was employed for sub-cellular resolution examination of both sliced and flat-mounted retinas. Two-photon imaging of retinal slices revealed autofluorescence signals over multiple functional layers, including the photoreceptor layer (PRL), outer nuclear layer (ONL), outer plexiform layer (OPL), inner nuclear layer (INL), inner plexiform layer (IPL), and ganglion cell layer (GCL). Using flat-mounted retinas, depth-resolved imaging of individual retinal layers further confirmed multiple sources of autofluorescence signals. Cellular structures were clearly observed at the PRL, ONL, INL, and GCL. At the PRL, the autofluorescence was dominantly recorded from the intracellular compartment of the photoreceptors; while mixed intracellular and extracellular autofluorescence signals were observed at the ONL, INL, and GCL. High resolution autofluorescence imaging clearly revealed mosaic organization of rod and cone photoreceptors; and sub-cellular bright autofluorescence spots, which might relate to connecting cilium, was observed in the cone photoreceptors only. Moreover, single-cone and double-cone outer segments could be directly differentiated.

Keywords: (170.2655) Functional monitoring and imaging; (170.3880) Medical and biological imaging; (170.4580) Optical diagnostics for medicine; (330.5380) Physiology

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