Noninvasive Two-Photon Microscopy Imaging of Mouse Retina and Retinal Pigment Epithelium

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2018 Oct 17

PMID 30324453

Citations 7

Authors

Grazyna Palczewska

Timothy S Kern

Krzysztof Palczewski

Affiliations

Soon will be listed here.

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.

Citing Articles

Safer and efficient base editing and prime editing via ribonucleoproteins delivered through optimized lipid-nanoparticle formulations.

Holubowicz R, Du S, Felgner J, Smidak R, Choi E, Palczewska G Nat Biomed Eng. 2024; 9(1):57-78.

PMID: 39609561 PMC: 11754100. DOI: 10.1038/s41551-024-01296-2.

Fiber optic-based integrated system for multiscale pharmacokinetic monitoring.

Li Y, Yang Y, Li W, Chen C, Lin Q, Huang H Biomed Opt Express. 2024; 15(6):3770-3782.

PMID: 38867773 PMC: 11166437. DOI: 10.1364/BOE.523179.

Lipofuscin, Its Origin, Properties, and Contribution to Retinal Fluorescence as a Potential Biomarker of Oxidative Damage to the Retina.

Rozanowska M Antioxidants (Basel). 2023; 12(12).

PMID: 38136230 PMC: 10740933. DOI: 10.3390/antiox12122111.

Distinct mouse models of Stargardt disease display differences in pharmacological targeting of ceramides and inflammatory responses.

Engfer Z, Lewandowski D, Dong Z, Palczewska G, Zhang J, Kordecka K Proc Natl Acad Sci U S A. 2023; 120(50):e2314698120.

PMID: 38064509 PMC: 10723050. DOI: 10.1073/pnas.2314698120.

Detailed 3D micro-modeling of rat aqueous drainage channels based on two-photon imaging: simulating aqueous humor through trabecular meshwork and Schlemm's canal by two-way fluid structure interaction approach.

Zhang J, Qian X, Zhang H, Chu H, Xu H, Liu Z Med Biol Eng Comput. 2022; 60(7):1915-1927.

PMID: 35524088 DOI: 10.1007/s11517-022-02580-6.

References

Imanishi Y, Gerke V, Palczewski K . Retinosomes: new insights into intracellular managing of hydrophobic substances in lipid bodies. J Cell Biol. 2004; 166(4):447-53. PMC: 1360213. DOI: 10.1083/jcb.200405110. View

Palczewska G, Maeda T, Imanishi Y, Sun W, Chen Y, Williams D . Noninvasive multiphoton fluorescence microscopy resolves retinol and retinal condensation products in mouse eyes. Nat Med. 2010; 16(12):1444-9. PMC: 3057900. DOI: 10.1038/nm.2260. View

Sharma R, Yin L, Geng Y, Merigan W, Palczewska G, Palczewski K . In vivo two-photon imaging of the mouse retina. Biomed Opt Express. 2013; 4(8):1285-93. PMC: 3756587. DOI: 10.1364/BOE.4.001285. View

Denk W, Strickler J, Webb W . Two-photon laser scanning fluorescence microscopy. Science. 1990; 248(4951):73-6. DOI: 10.1126/science.2321027. View

Tsina E, Chen C, Koutalos Y, Ala-Laurila P, Tsacopoulos M, Wiggert B . Physiological and microfluorometric studies of reduction and clearance of retinal in bleached rod photoreceptors. J Gen Physiol. 2004; 124(4):429-43. PMC: 2233903. DOI: 10.1085/jgp.200409078. View

Maeda A, Palczewska G, Golczak M, Kohno H, Dong Z, Maeda T . Two-photon microscopy reveals early rod photoreceptor cell damage in light-exposed mutant mice. Proc Natl Acad Sci U S A. 2014; 111(14):E1428-37. PMC: 3986171. DOI: 10.1073/pnas.1317986111. View

Thibos L, Applegate R, Schwiegerling J, Webb R . Standards for reporting the optical aberrations of eyes. J Refract Surg. 2002; 18(5):S652-60. DOI: 10.3928/1081-597X-20020901-30. View

Pawlicki M, Collins H, Denning R, Anderson H . Two-photon absorption and the design of two-photon dyes. Angew Chem Int Ed Engl. 2009; 48(18):3244-66. DOI: 10.1002/anie.200805257. View

Cua M, Wahl D, Zhao Y, Lee S, Bonora S, Zawadzki R . Coherence-Gated Sensorless Adaptive Optics Multiphoton Retinal Imaging. Sci Rep. 2016; 6:32223. PMC: 5013266. DOI: 10.1038/srep32223. View

10.

Palczewska G, Vinberg F, Stremplewski P, Bircher M, Salom D, Komar K . Human infrared vision is triggered by two-photon chromophore isomerization. Proc Natl Acad Sci U S A. 2014; 111(50):E5445-54. PMC: 4273384. DOI: 10.1073/pnas.1410162111. View

11.

Palczewska G, Dong Z, Golczak M, Hunter J, Williams D, Alexander N . Noninvasive two-photon microscopy imaging of mouse retina and retinal pigment epithelium through the pupil of the eye. Nat Med. 2014; 20(7):785-9. PMC: 4087080. DOI: 10.1038/nm.3590. View

12.

Palczewska G, Maeda A, Golczak M, Arai E, Dong Z, Perusek L . Receptor MER Tyrosine Kinase Proto-oncogene (MERTK) Is Not Required for Transfer of Bis-retinoids to the Retinal Pigmented Epithelium. J Biol Chem. 2016; 291(52):26937-26949. PMC: 5207201. DOI: 10.1074/jbc.M116.764563. View

13.

Kiser P, Palczewski K . Retinoids and Retinal Diseases. Annu Rev Vis Sci. 2016; 2:197-234. PMC: 5132409. DOI: 10.1146/annurev-vision-111815-114407. View

14.

Zipfel W, Williams R, Christie R, Nikitin A, Hyman B, Webb W . Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation. Proc Natl Acad Sci U S A. 2003; 100(12):7075-80. PMC: 165832. DOI: 10.1073/pnas.0832308100. View

15.

Imanishi Y, Batten M, Piston D, Baehr W, Palczewski K . Noninvasive two-photon imaging reveals retinyl ester storage structures in the eye. J Cell Biol. 2004; 164(3):373-83. PMC: 1360214. DOI: 10.1083/jcb.200311079. View

16.

Zwick H, Edsall P, Stuck B, Wood E, Elliott R, Cheramie R . Laser induced photoreceptor damage and recovery in the high numerical aperture eye of the garter snake. Vision Res. 2008; 48(3):486-93. DOI: 10.1016/j.visres.2007.09.012. View

17.

Stremplewski P, Komar K, Palczewski K, Wojtkowski M, Palczewska G . Periscope for noninvasive two-photon imaging of murine retina in vivo. Biomed Opt Express. 2015; 6(9):3352-61. PMC: 4574663. DOI: 10.1364/BOE.6.003352. View