Abstract
Two-photon excitation microscopy is perfectly suited for imaging deep into the retina due to its use of infrared (IR) wavelengths to excite endogenous fluorophores such as vitamin A-derived retinoids present in this tissue. Furthermore, two-photon excitation occurs only around a small focal volume, and scattered IR photons cannot excite retinal chromophores. These characteristics contribute to subcellular resolution and low noise of images obtained from deep within retinal layers. Here we describe how to customize a two-photon microscope for noninvasive imaging of the retina and retinal pigment epithelium (RPE) in the mouse eye, along with detailed instructions for mouse handling and retinal imaging, and we provide examples of mouse retinal two-photon microscopy data.
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Acknowledgments
We thank Dr. Leslie T. Webster, Jr., and members of the Palczewski laboratory for valuable comments regarding this manuscript. K.P. is chief scientific officer at Polgenix. K.P. is an inventor of the US patent no. 7,706,863 and US patent no. 8,346,345, whose values may be affected by this publication. This research was supported in part by grants from the National Institutes of Health (NIH) (EY027283, EY025451, and EY024864).
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Palczewska, G., Kern, T.S., Palczewski, K. (2019). Noninvasive Two-Photon Microscopy Imaging of Mouse Retina and Retinal Pigment Epithelium. In: Weber, B.H.F., Langmann, T. (eds) Retinal Degeneration. Methods in Molecular Biology, vol 1834. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8669-9_21
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DOI: https://doi.org/10.1007/978-1-4939-8669-9_21
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