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Noninvasive two-photon microscopy imaging of mouse retina and retinal pigment epithelium through the pupil of the eye

2014; Nature Portfolio; Volume: 20; Issue: 7 Linguagem: Inglês

10.1038/nm.3590

1546-170X

Grażyna Palczewska, Zhiqian Dong, Marcin Golczak, Jennifer J. Hunter, David R. Williams, Nathan Alexander, Krzysztof Palczewski,

Retinal Diseases and Treatments

Repetitive dynamic two-photon imaging of retinoid cycle fluorophores and subcellular details of their location within the retinal pigment epithelium in intact eyes of live mice. Two-photon excitation microscopy can image retinal molecular processes in vivo. Intrinsically fluorescent retinyl esters in subcellular structures called retinosomes are an integral part of the visual chromophore regeneration pathway. Fluorescent condensation products of all-trans-retinal accumulate in the eye with age and are also associated with age-related macular degeneration (AMD). Here, we report repetitive, dynamic imaging of these compounds in live mice through the pupil of the eye. By leveraging advanced adaptive optics, we developed a data acquisition algorithm that permitted the identification of retinosomes and condensation products in the retinal pigment epithelium by their characteristic localization, spectral properties and absence in genetically modified or drug-treated mice. This imaging approach has the potential to detect early molecular changes in retinoid metabolism that trigger light- and AMD-induced retinal defects and to assess the effectiveness of treatments for these conditions.