Near Infrared Autofluorescence Lifetime Imaging of Human Retinal Pigment Epithelium Using Adaptive Optics Scanning Light Ophthalmoscopy

Overview

Journal Invest Ophthalmol Vis Sci

Specialty Ophthalmology

Date 2024 May 17

PMID 38758638

Authors

Karteek Kunala

Janet A H Tang

Kristen E Bowles Johnson

Khang T Huynh

Keith Parkins

Hye-Jin Kim

Qiang Yang

Janet R Sparrow

Jennifer J Hunter

Affiliations

Soon will be listed here.

Abstract

Purpose: To demonstrate the first near-infrared adaptive optics fluorescence lifetime imaging ophthalmoscopy (NIR-AOFLIO) measurements in vivo of the human retinal pigment epithelial (RPE) cellular mosaic and to visualize lifetime changes at different retinal eccentricities.

Methods: NIR reflectance and autofluorescence were captured using a custom adaptive optics scanning light ophthalmoscope in 10 healthy subjects (23-64 years old) at seven eccentricities and in two eyes with retinal abnormalities. Repeatability was assessed across two visits up to 8 weeks apart. Endogenous retinal fluorophores and hydrophobic whole retinal extracts of Abca4-/- pigmented and albino mice were imaged to probe the fluorescence origin of NIR-AOFLIO.

Results: The RPE mosaic was resolved at all locations in five of seven younger subjects (<35 years old). The mean lifetime across near-peripheral regions (8° and 12°) was longer compared to near-foveal regions (0° and 2°). Repeatability across two visits showed moderate to excellent correlation (intraclass correlation: 0.88 [τm], 0.75 [τ1], 0.65 [τ2], 0.98 [a1]). The mean lifetime across drusen-containing eyes was longer than in age-matched healthy eyes. Fluorescence was observed in only the extracts from pigmented Abca4-/- mouse.

Conclusions: NIR-AOFLIO was repeatable and allowed visualization of the RPE cellular mosaic. An observed signal in only the pigmented mouse extract infers the fluorescence signal originates predominantly from melanin. Variations observed across the retina with intermediate age-related macular degeneration suggest NIR-AOFLIO may act as a functional measure of a biomarker for in vivo monitoring of early alterations in retinal health.

Citing Articles

From Cellular to Metabolic: Advances in Imaging of Inherited Retinal Diseases.

Parameswarappa D, Kulkarni A, Sahoo N, Padhy S, Singh S, Heon E Diagnostics (Basel). 2025; 15(1.

PMID: 39795556 PMC: 11720060. DOI: 10.3390/diagnostics15010028.

Near-Infrared Autofluorescence: Early Detection of Retinal Pigment Epithelial Alterations in Inherited Retinal Dystrophies.

Kellner S, Weinitz S, Farmand G, Kellner U J Clin Med. 2024; 13(22).

PMID: 39598030 PMC: 11594703. DOI: 10.3390/jcm13226886.

Multispectral label-free cellular imaging of human retinal pigment epithelium using adaptive optics fluorescence lifetime ophthalmoscopy improves feasibility for low emission analysis and increases sensitivity for detecting changes with age and....

Kunala K, Tang J, Parkins K, Hunter J J Biomed Opt. 2024; 29(Suppl 2):S22707.

PMID: 38962492 PMC: 11221116. DOI: 10.1117/1.JBO.29.S2.S22707.

References

Dysli C, Wolf S, Hatz K, Zinkernagel M . Fluorescence Lifetime Imaging in Stargardt Disease: Potential Marker for Disease Progression. Invest Ophthalmol Vis Sci. 2016; 57(3):832-41. DOI: 10.1167/iovs.15-18033. View

Huynh K, Walters S, Foley E, Hunter J . Separate lifetime signatures of macaque S cones, M/L cones, and rods observed with adaptive optics fluorescence lifetime ophthalmoscopy. Sci Rep. 2023; 13(1):2456. PMC: 9922306. DOI: 10.1038/s41598-023-28877-6. View

Palczewska G, Boguslawski J, Stremplewski P, Kornaszewski L, Zhang J, Dong Z . Noninvasive two-photon optical biopsy of retinal fluorophores. Proc Natl Acad Sci U S A. 2020; 117(36):22532-22543. PMC: 7486747. DOI: 10.1073/pnas.2007527117. View

Durairaj C, Chastain J, Kompella U . Intraocular distribution of melanin in human, monkey, rabbit, minipig and dog eyes. Exp Eye Res. 2012; 98:23-7. PMC: 3358346. DOI: 10.1016/j.exer.2012.03.004. View

Grieve K, Gofas-Salas E, Ferguson R, Sahel J, Paques M, Rossi E . near-infrared autofluorescence imaging of retinal pigment epithelial cells with 757 nm excitation. Biomed Opt Express. 2019; 9(12):5946-5961. PMC: 6490976. DOI: 10.1364/BOE.9.005946. View

Duncker T, Marsiglia M, Lee W, Zernant J, Tsang S, Allikmets R . Correlations among near-infrared and short-wavelength autofluorescence and spectral-domain optical coherence tomography in recessive Stargardt disease. Invest Ophthalmol Vis Sci. 2014; 55(12):8134-43. PMC: 4266077. DOI: 10.1167/iovs.14-14848. View

Ehlers A, Riemann I, Stark M, Konig K . Multiphoton fluorescence lifetime imaging of human hair. Microsc Res Tech. 2006; 70(2):154-61. DOI: 10.1002/jemt.20395. View

Weiter J, Delori F, Wing G, FITCH K . Retinal pigment epithelial lipofuscin and melanin and choroidal melanin in human eyes. Invest Ophthalmol Vis Sci. 1986; 27(2):145-52. View

Rozanowski B, Burke J, Boulton M, Sarna T, Rozanowska M . Human RPE melanosomes protect from photosensitized and iron-mediated oxidation but become pro-oxidant in the presence of iron upon photodegradation. Invest Ophthalmol Vis Sci. 2008; 49(7):2838-47. DOI: 10.1167/iovs.08-1700. View

10.

N Keilhauer C, Delori F . Near-infrared autofluorescence imaging of the fundus: visualization of ocular melanin. Invest Ophthalmol Vis Sci. 2006; 47(8):3556-64. DOI: 10.1167/iovs.06-0122. View

11.

Cideciyan A, Swider M, Aleman T, Roman M, Sumaroka A, Schwartz S . Reduced-illuminance autofluorescence imaging in ABCA4-associated retinal degenerations. J Opt Soc Am A Opt Image Sci Vis. 2007; 24(5):1457-67. PMC: 2579898. DOI: 10.1364/josaa.24.001457. View

12.

Sauer L, Vitale A, Milliken C, Modersitzki N, Blount J, Bernstein P . Autofluorescence Lifetimes Measured with Fluorescence Lifetime Imaging Ophthalmoscopy (FLIO) Are Affected by Age, but Not by Pigmentation or Gender. Transl Vis Sci Technol. 2020; 9(9):2. PMC: 7442880. DOI: 10.1167/tvst.9.9.2. View

13.

Konig K . Clinical multiphoton tomography. J Biophotonics. 2009; 1(1):13-23. DOI: 10.1002/jbio.200710022. View

14.

Deyl Z, Macek K, Adam M, Vancikova O . Studies on the chemical nature of elastin fluorescence. Biochim Biophys Acta. 1980; 625(2):248-54. DOI: 10.1016/0005-2795(80)90288-3. View

15.

Paavo M, Zhao J, Kim H, Lee W, Zernant J, Cai C . Mutations in GPR143/OA1 and ABCA4 Inform Interpretations of Short-Wavelength and Near-Infrared Fundus Autofluorescence. Invest Ophthalmol Vis Sci. 2018; 59(6):2459-2469. PMC: 5959512. DOI: 10.1167/iovs.18-24213. View

16.

Schwanengel L, Weber S, Simon R, Lehmann T, Augsten R, Meller D . Changes in drusen-associated autofluorescence over time observed by fluorescence lifetime imaging ophthalmoscopy in age-related macular degeneration. Acta Ophthalmol. 2022; 101(2):e154-e166. DOI: 10.1111/aos.15238. View

17.

Laforest T, Kunzi M, Kowalczuk L, Carpentras D, Behar-Cohen F, Moser C . Transscleral Optical Phase Imaging of the Human Retina. Nat Photonics. 2020; 14(7):439-445. PMC: 7326609. DOI: 10.1038/s41566-020-0608-y. View

18.

Ince C, Coremans J, Bruining H . In vivo NADH fluorescence. Adv Exp Med Biol. 1992; 317:277-96. DOI: 10.1007/978-1-4615-3428-0_30. View

19.

Tang J, Granger C, Kunala K, Parkins K, Huynh K, Bowles-Johnson K . Adaptive optics fluorescence lifetime imaging ophthalmoscopy of human retinal pigment epithelium. Biomed Opt Express. 2022; 13(3):1737-1754. PMC: 8973160. DOI: 10.1364/BOE.451628. View

20.

Greenstein V, Schuman A, Lee W, Duncker T, Zernant J, Allikmets R . Near-infrared autofluorescence: its relationship to short-wavelength autofluorescence and optical coherence tomography in recessive stargardt disease. Invest Ophthalmol Vis Sci. 2015; 56(5):3226-34. PMC: 4453463. DOI: 10.1167/iovs.14-16050. View