OSA's Digital Library

Journal of the Optical Society of America A

Journal of the Optical Society of America A


  • Vol. 19, Iss. 6 — Jun. 1, 2002
  • pp: 1172–1186

In vivo resonant Raman measurement of macular carotenoid pigments in the young and the aging human retina

Werner Gellermann, Igor V. Ermakov, Maia R. Ermakova, Robert W. McClane, Da-You Zhao, and Paul S. Bernstein  »View Author Affiliations

JOSA A, Vol. 19, Issue 6, pp. 1172-1186 (2002)

View Full Text Article

Acrobat PDF (1216 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



We have used resonant Raman scattering spectroscopy as a novel, noninvasive, in vivo optical technique to measure the concentration of the macular carotenoid pigments lutein and zeaxanthin in the living human retina of young and elderly adults. Using a backscattering geometry and resonant molecular excitation in the visible wavelength range, we measure the Raman signals originating from the single- and double-bond stretch vibrations of the π-conjugated molecule’s carbon backbone. The Raman signals scale linearly with carotenoid content, and the required laser excitation is well below safety limits for macular exposure. Furthermore, the signals decline significantly with increasing age in normal eyes. The Raman technique is objective and quantitative and may lead to a new method for rapid screening of carotenoid pigment levels in large populations at risk for vision loss from age-related macular degeneration, the leading cause of blindness in the elderly in the United States.

© 2002 Optical Society of America

OCIS Codes
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(170.1610) Medical optics and biotechnology : Clinical applications
(170.4470) Medical optics and biotechnology : Ophthalmology
(170.4580) Medical optics and biotechnology : Optical diagnostics for medicine
(300.6450) Spectroscopy : Spectroscopy, Raman
(330.4300) Vision, color, and visual optics : Vision system - noninvasive assessment

Werner Gellermann, Igor V. Ermakov, Maia R. Ermakova, Robert W. McClane, Da-You Zhao, and Paul S. Bernstein, "In vivo resonant Raman measurement of macular carotenoid pigments in the young and the aging human retina," J. Opt. Soc. Am. A 19, 1172-1186 (2002)

Sort:  Author  |  Year  |  Journal  |  Reset


  1. R. A. Bone, J. T. Landrum, and S. L. Tarsis, “Preliminary identification of the human macular pigment,” Vision Res. 25, 1531–1535 (1985).
  2. G. J. Handelman, D. M. Snodderly, A. J. Adler, M. D. Russett, and E. A. Dratz, “Measurement of carotenoids in human and monkey retinas,” Methods Enzymol. 213, 220–230 (1992).
  3. W. Schalch, P. Dayhaw-Barker, and F. M. Barker, “The carotenoids of the human retina,” in Nutritional and Environmental Influences on the Eye, A. Taylor, ed. (CRC, Boca Raton, Fla., 1999), pp. 215–250.
  4. D. M. Snodderly, “Evidence for protection against age-related macular degeneration by carotenoids and antioxidant vitamins,” Am. J. Clin. Nutr. Suppl. 62, 1448S–1461S (1995).
  5. Eye Disease Case Control Study Group, “Antioxidant status and neovascular age-related macular degeneration,” Arch. Ophthalmol. (Chicago) 111, 104–109 (1993).
  6. J. M. Seddon, U. A. Ajani, R. D. Sperduto, R. Hiller, N. Blair, T. C. Burton, M. D. Farber, E. S. Gragoudas, J. Haller, D. T. Miller, L. A. Yannuzzi, and W. Willet, “Dietary carotenoids, vitamins A, C, and E, and advanced age-related macular degeneration,” J. Am. Med. Assoc. 272, 1413–1420 (1994).
  7. B. R. Hammond, E. J. Johnson, R. M. Russell, N. I. Krinsky, K.-J. Yeum, R. B. Edwards, and D. M. Snodderly, “Dietary modification of human macular pigment density,” Invest. Ophthalmol. Visual Sci. 38, 1795–1801 (1997).
  8. J. T. Landrum, R. A. Bone, H. Joa, M. D. Kilburn, L. L. Moore, and K. E. Sprague, “A one year study of the macular pigment: the effect of 140 days of a lutein supplement,” Exp. Eye Res. 65, 57–62 (1997).
  9. J. T. Landrum, R. A. Bone, and M. D. Kilburn, “The macular pigment: a possible role in protection from age-related macular degeneration,” Adv. Pharmacol. 38, 537–556 (1997).
  10. R. A. Bone, J. T. Landrum, S. T. Mayne, C. M. Gomez, S. E. Tibor, and E. E. Twaroska, “Macular pigment in donor eyes with and without AMD: a case-control study,” Invest. Ophthalmol. Visual Sci. 42, 235–240 (2000).
  11. Age Related Eye Disease Study Research Group, “A randomized, placebo controlled clinical trial of high-dose supplementation with vitamins C and E, beta carotene, and zinc for age-related macular degeneration and vision loss,” Arch. Ophthalmol. 119, 1417–1436 (2001).
  12. D. M. Snodderly, J. D. Auran, and F. C. Delori, “The macular pigment, I: absorbance spectra, localization, and discrimination from other yellow pigments in primate retinas,” Invest. Ophthalmol. Visual Sci. 25, 660–673 (1984).
  13. R. A. Bone, J. T. Landrum, L. Fernandez, and S. L. Tarsis, “Analysis of macular pigment by HPLC: retinal distribution and age study,” Invest. Ophthalmol. Visual Sci. 29, 843–849 (1988).
  14. R. A. Bone, J. T. Landrum, L. M. Friedes, C. A. Gomez, M. A. Kilburn, E. Menendez, I. Vidal, and W. Wang, “Distribution of lutein and zeaxanthin stereoisomers in the human retina,” Exp. Eye Res. 64, 211–218 (1997).
  15. D. M. Snodderly, G. J. Handelman, and A. A. Adler, “Distribution of individual macular pigment carotenoids in central retina of macaque and squirrel monkeys,” Invest. Ophthalmol. Visual Sci. 32, 268–279 (1991).
  16. J. D. M. Gass, “The Müller cell cone, an overlooked part of the anatomy of the fovea centralis,” Arch. Ophthalmol. 117, 821–823 (1999).
  17. O. G. Sommerberg, W. G. Siems, J. S. Hurst, J. W. Lewis, D. S. Kliger, and F. J. van Kuijk, “Lutein and zeaxanthin are associated with photoreceptors in the human retina,” Curr. Eye Res. 19, 502–505 (1999).
  18. L. M. Rapp, S. S. Maple, and J. H. Choi, “Lutein and zeaxanthin concentrations in rod outer segment membranes from perifoveal and peripheral human retina,” Invest. Ophthalmol. Visual Sci. 41, 1200–1209 (2000).
  19. B. R. Hammond, Jr., B. R. Wooten, and D. M. Snodderly, “Individual variations in the spatial profile of human macular pigment,” J. Opt. Soc. Am. A 14, 1187–1196 (1997).
  20. P. S. Bernstein and N. B. Katz, “The role of ocular free radicals in age-related macular degeneration,” in Environmental Stressors: Effects on Lung, Skin, Eye and Immune System Function, J. Fuchs and L. Packer, eds. (Marcel Dekker, New York, 2000), pp. 423–456.
  21. S. Beatty, H.-H. Koh, D. Henson, and M. Boulton, “The role of oxidative stress in the pathogenesis of age-related macular degeneration,” Surv. Ophthalmol. 45, 115–134 (2000).
  22. V. M. Reading and R. A. Weale, “Macular pigment and chromatic aberration,” J. Am. Optom. Assoc. 64, 231–234 (1974).
  23. S. Beatty, I. J. Murray, D. B. Henson, D. Carden, H.-H. Koh, and M. E. Boulton, “Macular pigment and risk for age-related macular degeneration in subjects from a northern European population,” Invest. Ophthalmol. Visual Sci. 42, 439–446 (2001).
  24. D. M. Snodderly and B. R. Hammond, “In vivo psychophysical assessment of nutritional and environmental influences on human ocular tissues: lens and macular pigment,” in Nutritional and Environmental Influences on the Eye, A. Taylor, ed. (CRC, Boca Raton, Fla., 1999), pp. 251–273.
  25. J. S. Werner, M. L. Bieber, and B. E. Schefrin, “Senescence of foveal and parafoveal cone sensitivities and their relations to macular pigment density,” J. Opt. Soc. Am. A 17, 1918–1932 (2000).
  26. B. R. Hammond and M. Caruso-Avery, “Macular pigment optical density in a southwestern sample,” Invest. Ophthalmol. Visual Sci. 41, 1492–1497 (2000).
  27. D. van Norren and L. F. Tiemeijer, “Spectral reflectance of the human eye,” Vision Res. 26, 313–320 (1986).
  28. F. C. Delori and K. P. Pflibsen, “Spectral reflectance of the human ocular fundus,” Appl. Opt. 28, 1061–1077 (1989).
  29. J. van de Kraats, T. T. J. M. Berendshot, and D. van Norren, “The pathways of light measured in fundus reflectometry,” Vision Res. 36, 2229–2247 (1996).
  30. T. Berendshot, J. van de Kraats, and D. van Norren, “Three methods to measure macular pigment compared in a lutein intake study,” Invest. Ophthalmol. Visual Sci. Suppl. 39, S314 (1999).
  31. P. E. Kilbride, K. R. Alexander, M. Fishman, and G. A. Fishman, “Human macular pigment assessed by imaging fundus reflectometry,” Vision Res. 29, 663–674 (1989).
  32. R. H. Webb, G. W. Hughes, and O. Pomerantzeff, “Flying spot TV ophthalmoscope,” Appl. Opt. 19, 2991–2997 (1980).
  33. R. W. Webb, G. W. Hughes, and F. C. Delori, “Confocal scanning laser ophthalmosope,” Appl. Opt. 26, 1492–1499 (1987).
  34. D. van Norren and J. van de Kraats, “Imaging retinal densitometry with a confocal scanning ophthalmoscope,” Vision Res. 29, 1825–1830 (1989).
  35. A. E. Elsner, S. A. Burns, F. C. Delori, and R. H. Webb, “Quantitative reflectometry with the SLO,” in Laser Scanning Ophthalmoscopy and Tomography, J. E. Nasemann and R. O. W. Burk, eds. (Quintessenz-Verlag, Berlin, 1990), pp. 109–121.
  36. A. E. Elsner, S. A. Burns, G. W. Hughes, and R. H. Webb, “Reflectometry with a scanning laser ophthalmoscope,” Appl. Opt. 31, 3697–3710 (1992).
  37. A. E. Elsner, S. A. Burns, E. Beausencourt, and J. J. Weiter, “Foveal cone photopigment distribution: small alterations associated with macular pigment distribution,” Invest. Ophthalmol. Visual Sci. 39, 2394–2404 (1998).
  38. D. Schweitzer, M. Hammer, and M. Scibor, “Imaging spectrometry in ophthalmology: principle and applications in microcirculation and in investigation of pigments,” Invest. Ophthalmol. Visual Sci. 39, 2001–2011 (1998).
  39. F. C. Delori, “Macular pigment density measured by reflectometry and fluorophotometry,” in Ophthalmic and Visual Optics and Noninvasive Assessment of the Visual System, Vol. 3 of 1993 OSA Technical Digest Series (Optical Society of America, Washington, D.C., 1993), pp. 240–243.
  40. F. C. Delori, D. G. Goger, B. R. Hammond, D. M. Snodderly, and S. A. Burns, “Macular pigment density measured by autofluorescence spectrometry: comparison with reflectometry and heterochromatic flicker photometry,” J. Opt. Soc. Am. A 18, 1212–1230 (2001).
  41. T. T. J. M. Berendshot, R. A. Goldbohm, W. A. A. Kloepping, J. van de Kraats, J. Van Norel, and D. van Norren, “Influence of lutein supplementation on macular pigment, assessed with two objective techniques,” Invest. Ophthalmol. Visual Sci. 41, 3322–3326 (2000).
  42. W. Gellermann, M. D. Yoshida, R. W. McClane, N. Balashov, and P. S. Bernstein, “Raman detection of pigments in the human retina,” in Optical Biopsy II, R. R. Alfano and A. Katzir, eds., Proc. SPIE 3250, 8–17 (1998).
  43. P. S. Bernstein, M. D. Yoshida, N. B. Katz, R. W. McClane, and W. Gellermann, “Raman detection of macular carotenoid pigments in intact human retina,” Invest. Ophthalmol. Visual Sci. 39, 2003–2011 (1998).
  44. I. V. Ermakov, R. W. McClane, W. Gellermann, and P. S. Bernstein, “Resonant Raman detection of macular pigment levels in the living human retina,” Opt. Lett. 26, 202–204 (2001).
  45. P. S. Bernstein, W. Gellermann, and R. W. McClane, “Measurement of macular carotenoid levels in the human retina using Raman spectroscopy,” U.S. patent 5,873,831, February 23, 1999.
  46. W. Gellermann, R. W. McClane, N. B. Katz, and P. S. Bernstein, “Method and apparatus for noninvasive measurement of carotenoids and related chemical substances in biological tissue,” U.S. patent 6,205,354, March 20, 2001.
  47. T. R. Hata, T. A. Scholz, I. V. Ermakov, R. W. McClane, F. Khachik, W. Gellermann, and L. K. Pershing, “Non-invasive Raman spectroscopic detection of carotenoids in human skin,” J. Invest. Dermatol. 115, 441–448 (2000).
  48. A. P. Shreve, J. K. Trautman, T. G. Owens, and A. C. Albrecht, “Determination of the S2 lifetime of β-carotene,” Chem. Phys. Lett. 178, 89 (1991).
  49. Y. Koyama, I. Takatsuka, M. Nakata, and M. Tasumi, “Raman and infrared spectra of the all-trans, 7-cis, 9-cis, 13-cis, and 15-cis isomers of β-carotene: key bands distinguishing stretched or terminal-bent configurations from central-bent configurations” J. Raman Spectrosc. 19, 37–49 (1988).
  50. A. C. Albrecht, “On the theory of Raman intensities,” J. Chem. Phys. 34, 1476–1484 (1961).
  51. W. Kiefer and M. Spiekermann, “Resonance Raman theories,” in Infrared and Raman Spectra, B. Schrader, ed. (VCH Verlagsgesellschaft, Weinheim, Germany, 1995), pp. 468–473.
  52. F. C. Delori, J. S. Parker, and M. A. Mainster, “Light levels in fundus photography and fluorescein angiography,” Vision Res. 20, 1099–1104 (1980).
  53. American National Standards Institute, “American national standard for safe use of lasers,” ANSI Z136.1-2000 (Laser Institute of America, Orlando, Fla., 2000), Sec. 8.3.
  54. L. Apt and A. Henrick, “Pupillary dilatation with single eyedrop mydriatic combinations,” Am. J. Ophthalmol. 89, 553–559 (1980).
  55. C. J. Hammond, H. Sneider, T. D. Spector, and C. E. Gilbert, “Factors affecting pupil size after dilatation: the Twin Eye Study,” Br. J. Ophthamol. 84, 1173–1176 (2000).
  56. P. S. Bernstein, F. Khachik, L. S. Carvalho, G. J. Muir, D. Y. Zhao, and N. B. Katz, “Identification and quantitation of carotenoids and their metabolites in the tissues of the human eye,” Exp. Eye Res. 72, 215–223 (2001).
  57. E. A. Boettner and J. R. Wolter, “Transmission of the ocular media,” Invest. Ophthalmol. Visual Sci. 1, 776–783 (1962).
  58. P. A. Sample, F. D. Esterson, R. N. Weinreb, and R. M. Boynton, “The aging lens: in vivo assessment of light absorption in 84 human eyes,” Invest. Ophthalmol. Visual Sci. 29, 1306–1311 (1988).
  59. J. Sebag, R. R. Ansari, S. Dunker, and K. I. Suh, “Dynamic light scattering of diabetic vitreopathy,” Diabetes Technol. Ther. 1, 169–176 (1999).
  60. J. Dillon, L. Zheng, J. C. Merriam, and E. R. Gaillard, “Transmission spectra of light to the mammalian retina,” Photochem. Photobiol. 71, 225–229 (2000).
  61. E. R. Gaillard, L. Zheng, J. C. Merriam, and J. Dillon, “Age-related changes in the absorption characteristics of the primate lens,” Invest. Ophthalmol. Visual Sci. 41, 1454–1459 (2000).
  62. R. A. Weale, “Age and the transmittance of the human crystalline lens,” J. Physiol. 395, 577–587 (1988).
  63. J. Pokorny, V. C. Smith, and M. Lutze, “Aging of the human lens,” Appl. Opt. 26, 1437–1440 (1987).
  64. A. E. Elsner, S. A. Burns, J. J. Weiter, and F. C. Delori, “Infrared imaging of subretinal structures in the human ocular fundus,” Vision Res. 36, 191–201 (1996).
  65. B. R. Hammond, B. R. Wooten, and D. M. Snodderly, “Preservation of visual sensitivity of older subjects: association with macular pigment density,” Invest. Ophthalmol. Visual Sci. 39, 397–406 (1998).

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited