Abstract

This work describes the selective targeting of pigmented retinal pigment epithelial (RPE) cells by a single pulsed laser irradiation. We observed: (1) single pulsed laser irradiation caused cellular damages on pigmented, and not on non-pigmented RPE cells at laser radiant exposure up to 2550 mJ/cm2; (2) in the mixture of pigmented and non-pigmented RPE cells, single pulsed laser-induced damage was confined to pigmented RPE cells. This study demonstrates that the pigmented RPE cells can be selectively damaged, using a single pulsed laser irradiation, without thermal coagulation to adjacent non-pigmented RPE cells.

© 2008 Optical Society of America

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  1. M. F. Marmor and T. J. Wolfensberger, The retinal pigment epithelium (Oxford University Press, New York, 1998).
  2. R. R. Anderson and J. R. Parrish, "Selective photothermolysis: precise microsurgery by selective absorption of pulsed radiation," Science 220, 524-527 (1983).
    [CrossRef] [PubMed]
  3. V. P. Gabel, R. Birngruber, and F. Hillenkamp, Visible and near infrared light absorption in pigment epithelium and choroids. In International Congress Series No.450, XXIII concilium Ophthalmologicum, K. Shimizu, ed.Princeton, NJ: Excerpta Medica:1978:658-662). (Excerpta Medica, Princeton, NJ, 1978).
    [PubMed]
  4. A. Leuenberger, "Treatment of diabetic retinopathy with photocoagulation," Mod. Probl. Ophthalmol. 10, 564-576 (1972).
    [PubMed]
  5. J. Roider, F. Hillenkamp, T. J. Flotte, and R. Birngruber, "Microphotocoagulation: Selective effects of repetitive short laser pulses," Proc. Natl. Acad. Sci. U.S.A. 90, 8643-8647 (1993).
    [CrossRef] [PubMed]
  6. R. Brinkmann, N. Koop, M. Oezdemir, C. Alt, G. Schuele, C. P. Lin, and R. Birngruber, "Targeting of the retinal pigment epithelium (RPE) by means of a rapidly scanned continuous wave (CW) laser beam," Lasers Surg. Med. 32, 252-264 (2003).
    [CrossRef] [PubMed]
  7. K. C. Dunn, A. E. Aotaki-Keen, F. R. Putkey, and L. M. Hjelmeland, "ARPE-19, A human retinal pigment epitheial cell line with differentiated properties," Exp. Eye Res. 62, 155-169 (1996).
    [CrossRef] [PubMed]
  8. B. L. Seagle, E. M. Gasyna, W. F. Mieler, and J. R. J. Norris, "Photoprotection of human retinal pigment epithelium cells against blue light-induced apoptosis by melanin free radicals from Sepia officinalis," Proc. Natl. Acad. Sci. U. S. A. 103, 16644-16648 (2006).
    [CrossRef] [PubMed]
  9. M. Saham, K. U. Loeffler, P. Seifer, and M. Spitznas, "Effect of ultraviolet radiation on melanogenesis in four different types of cultured bovine ocular pigmented cells," Graefe's Arch. Clin. Exp. Ophthalmol. 239, 302-309 (2001).
    [CrossRef]
  10. A. E. Siegman, Defining and Measuring Laser Beam Parameters. In Laser Beam Characterization (Sociedad Espanola de Optica, Madrid, 1993).
  11. S. Proulx, S. L. Guerin, and C. Salesse, "Effect of quiescence on integrin alpha5beta1 expression in human retinal pigment epithelium," Mol. Vis. 3, 473-481 (2003).
  12. R. Birngruber, F. Hillenkamp, and V. P. Gabel, "Theoretical investigations of laser thermal retinal injury," Health Phys. 48, 781-796 (1985).
    [CrossRef] [PubMed]
  13. C. P. Lin and M. W. Kelly, "Ultrafast time-resolved imaging of stress transient and cavitation from short pulsed laser irradiated melanin particles," Proc. SPIE 2391A, 294-299 (1995).
    [CrossRef]
  14. H. Lee, C. Alt, C. M. Pitsillides, and C. P. Lin, "Optical detection of intracellular cavitation during selective laser targeting of the retinal pigment epithelium: dependence of cell death mechanism on pulse duration," J. Biomed. Opt. 12, 064034 (2007).
    [CrossRef]
  15. R. Brinkmann, G. Hüttmann, J. Rögener, J. Roider, R. Birngruber, and C. P. Lin, "Origin of RPE-cell damage by pulsed laser irradiance in the ns to µs time regime," Lasers Surg. Med. 27, 451-464 (2000).
    [CrossRef] [PubMed]
  16. S. Y. Schmidt and R. D. Peisch, "Melanin concentration in normal human retinal pigment epithelium," Invest. Ophthalmol. Vis. Sci. 27, 1063-1067 (1986).
    [PubMed]
  17. R. Brinkmann, J. Roider, and R. Birngruber, "Selective retina therapy (SRT): a review on methods, techniques, preclinical and first clinical results," Bull. Soc. Belge. Ophtalmol. 302, 51-69 (2006).

2007

H. Lee, C. Alt, C. M. Pitsillides, and C. P. Lin, "Optical detection of intracellular cavitation during selective laser targeting of the retinal pigment epithelium: dependence of cell death mechanism on pulse duration," J. Biomed. Opt. 12, 064034 (2007).
[CrossRef]

2006

R. Brinkmann, J. Roider, and R. Birngruber, "Selective retina therapy (SRT): a review on methods, techniques, preclinical and first clinical results," Bull. Soc. Belge. Ophtalmol. 302, 51-69 (2006).

B. L. Seagle, E. M. Gasyna, W. F. Mieler, and J. R. J. Norris, "Photoprotection of human retinal pigment epithelium cells against blue light-induced apoptosis by melanin free radicals from Sepia officinalis," Proc. Natl. Acad. Sci. U. S. A. 103, 16644-16648 (2006).
[CrossRef] [PubMed]

2003

S. Proulx, S. L. Guerin, and C. Salesse, "Effect of quiescence on integrin alpha5beta1 expression in human retinal pigment epithelium," Mol. Vis. 3, 473-481 (2003).

R. Brinkmann, N. Koop, M. Oezdemir, C. Alt, G. Schuele, C. P. Lin, and R. Birngruber, "Targeting of the retinal pigment epithelium (RPE) by means of a rapidly scanned continuous wave (CW) laser beam," Lasers Surg. Med. 32, 252-264 (2003).
[CrossRef] [PubMed]

2001

M. Saham, K. U. Loeffler, P. Seifer, and M. Spitznas, "Effect of ultraviolet radiation on melanogenesis in four different types of cultured bovine ocular pigmented cells," Graefe's Arch. Clin. Exp. Ophthalmol. 239, 302-309 (2001).
[CrossRef]

2000

R. Brinkmann, G. Hüttmann, J. Rögener, J. Roider, R. Birngruber, and C. P. Lin, "Origin of RPE-cell damage by pulsed laser irradiance in the ns to µs time regime," Lasers Surg. Med. 27, 451-464 (2000).
[CrossRef] [PubMed]

1996

K. C. Dunn, A. E. Aotaki-Keen, F. R. Putkey, and L. M. Hjelmeland, "ARPE-19, A human retinal pigment epitheial cell line with differentiated properties," Exp. Eye Res. 62, 155-169 (1996).
[CrossRef] [PubMed]

1995

C. P. Lin and M. W. Kelly, "Ultrafast time-resolved imaging of stress transient and cavitation from short pulsed laser irradiated melanin particles," Proc. SPIE 2391A, 294-299 (1995).
[CrossRef]

1993

J. Roider, F. Hillenkamp, T. J. Flotte, and R. Birngruber, "Microphotocoagulation: Selective effects of repetitive short laser pulses," Proc. Natl. Acad. Sci. U.S.A. 90, 8643-8647 (1993).
[CrossRef] [PubMed]

1986

S. Y. Schmidt and R. D. Peisch, "Melanin concentration in normal human retinal pigment epithelium," Invest. Ophthalmol. Vis. Sci. 27, 1063-1067 (1986).
[PubMed]

1985

R. Birngruber, F. Hillenkamp, and V. P. Gabel, "Theoretical investigations of laser thermal retinal injury," Health Phys. 48, 781-796 (1985).
[CrossRef] [PubMed]

1983

R. R. Anderson and J. R. Parrish, "Selective photothermolysis: precise microsurgery by selective absorption of pulsed radiation," Science 220, 524-527 (1983).
[CrossRef] [PubMed]

1972

A. Leuenberger, "Treatment of diabetic retinopathy with photocoagulation," Mod. Probl. Ophthalmol. 10, 564-576 (1972).
[PubMed]

Alt, C.

H. Lee, C. Alt, C. M. Pitsillides, and C. P. Lin, "Optical detection of intracellular cavitation during selective laser targeting of the retinal pigment epithelium: dependence of cell death mechanism on pulse duration," J. Biomed. Opt. 12, 064034 (2007).
[CrossRef]

R. Brinkmann, N. Koop, M. Oezdemir, C. Alt, G. Schuele, C. P. Lin, and R. Birngruber, "Targeting of the retinal pigment epithelium (RPE) by means of a rapidly scanned continuous wave (CW) laser beam," Lasers Surg. Med. 32, 252-264 (2003).
[CrossRef] [PubMed]

Anderson, R. R.

R. R. Anderson and J. R. Parrish, "Selective photothermolysis: precise microsurgery by selective absorption of pulsed radiation," Science 220, 524-527 (1983).
[CrossRef] [PubMed]

Aotaki-Keen, A. E.

K. C. Dunn, A. E. Aotaki-Keen, F. R. Putkey, and L. M. Hjelmeland, "ARPE-19, A human retinal pigment epitheial cell line with differentiated properties," Exp. Eye Res. 62, 155-169 (1996).
[CrossRef] [PubMed]

Birngruber, R.

R. Brinkmann, J. Roider, and R. Birngruber, "Selective retina therapy (SRT): a review on methods, techniques, preclinical and first clinical results," Bull. Soc. Belge. Ophtalmol. 302, 51-69 (2006).

R. Brinkmann, N. Koop, M. Oezdemir, C. Alt, G. Schuele, C. P. Lin, and R. Birngruber, "Targeting of the retinal pigment epithelium (RPE) by means of a rapidly scanned continuous wave (CW) laser beam," Lasers Surg. Med. 32, 252-264 (2003).
[CrossRef] [PubMed]

R. Brinkmann, G. Hüttmann, J. Rögener, J. Roider, R. Birngruber, and C. P. Lin, "Origin of RPE-cell damage by pulsed laser irradiance in the ns to µs time regime," Lasers Surg. Med. 27, 451-464 (2000).
[CrossRef] [PubMed]

J. Roider, F. Hillenkamp, T. J. Flotte, and R. Birngruber, "Microphotocoagulation: Selective effects of repetitive short laser pulses," Proc. Natl. Acad. Sci. U.S.A. 90, 8643-8647 (1993).
[CrossRef] [PubMed]

R. Birngruber, F. Hillenkamp, and V. P. Gabel, "Theoretical investigations of laser thermal retinal injury," Health Phys. 48, 781-796 (1985).
[CrossRef] [PubMed]

Brinkmann, R.

R. Brinkmann, J. Roider, and R. Birngruber, "Selective retina therapy (SRT): a review on methods, techniques, preclinical and first clinical results," Bull. Soc. Belge. Ophtalmol. 302, 51-69 (2006).

R. Brinkmann, N. Koop, M. Oezdemir, C. Alt, G. Schuele, C. P. Lin, and R. Birngruber, "Targeting of the retinal pigment epithelium (RPE) by means of a rapidly scanned continuous wave (CW) laser beam," Lasers Surg. Med. 32, 252-264 (2003).
[CrossRef] [PubMed]

R. Brinkmann, G. Hüttmann, J. Rögener, J. Roider, R. Birngruber, and C. P. Lin, "Origin of RPE-cell damage by pulsed laser irradiance in the ns to µs time regime," Lasers Surg. Med. 27, 451-464 (2000).
[CrossRef] [PubMed]

Dunn, K. C.

K. C. Dunn, A. E. Aotaki-Keen, F. R. Putkey, and L. M. Hjelmeland, "ARPE-19, A human retinal pigment epitheial cell line with differentiated properties," Exp. Eye Res. 62, 155-169 (1996).
[CrossRef] [PubMed]

Flotte, T. J.

J. Roider, F. Hillenkamp, T. J. Flotte, and R. Birngruber, "Microphotocoagulation: Selective effects of repetitive short laser pulses," Proc. Natl. Acad. Sci. U.S.A. 90, 8643-8647 (1993).
[CrossRef] [PubMed]

Gabel, V. P.

R. Birngruber, F. Hillenkamp, and V. P. Gabel, "Theoretical investigations of laser thermal retinal injury," Health Phys. 48, 781-796 (1985).
[CrossRef] [PubMed]

Gasyna, E. M.

B. L. Seagle, E. M. Gasyna, W. F. Mieler, and J. R. J. Norris, "Photoprotection of human retinal pigment epithelium cells against blue light-induced apoptosis by melanin free radicals from Sepia officinalis," Proc. Natl. Acad. Sci. U. S. A. 103, 16644-16648 (2006).
[CrossRef] [PubMed]

Guerin, S. L.

S. Proulx, S. L. Guerin, and C. Salesse, "Effect of quiescence on integrin alpha5beta1 expression in human retinal pigment epithelium," Mol. Vis. 3, 473-481 (2003).

Hillenkamp, F.

J. Roider, F. Hillenkamp, T. J. Flotte, and R. Birngruber, "Microphotocoagulation: Selective effects of repetitive short laser pulses," Proc. Natl. Acad. Sci. U.S.A. 90, 8643-8647 (1993).
[CrossRef] [PubMed]

R. Birngruber, F. Hillenkamp, and V. P. Gabel, "Theoretical investigations of laser thermal retinal injury," Health Phys. 48, 781-796 (1985).
[CrossRef] [PubMed]

Hjelmeland, L. M.

K. C. Dunn, A. E. Aotaki-Keen, F. R. Putkey, and L. M. Hjelmeland, "ARPE-19, A human retinal pigment epitheial cell line with differentiated properties," Exp. Eye Res. 62, 155-169 (1996).
[CrossRef] [PubMed]

Hüttmann, G.

R. Brinkmann, G. Hüttmann, J. Rögener, J. Roider, R. Birngruber, and C. P. Lin, "Origin of RPE-cell damage by pulsed laser irradiance in the ns to µs time regime," Lasers Surg. Med. 27, 451-464 (2000).
[CrossRef] [PubMed]

Kelly, M. W.

C. P. Lin and M. W. Kelly, "Ultrafast time-resolved imaging of stress transient and cavitation from short pulsed laser irradiated melanin particles," Proc. SPIE 2391A, 294-299 (1995).
[CrossRef]

Koop, N.

R. Brinkmann, N. Koop, M. Oezdemir, C. Alt, G. Schuele, C. P. Lin, and R. Birngruber, "Targeting of the retinal pigment epithelium (RPE) by means of a rapidly scanned continuous wave (CW) laser beam," Lasers Surg. Med. 32, 252-264 (2003).
[CrossRef] [PubMed]

Lee, H.

H. Lee, C. Alt, C. M. Pitsillides, and C. P. Lin, "Optical detection of intracellular cavitation during selective laser targeting of the retinal pigment epithelium: dependence of cell death mechanism on pulse duration," J. Biomed. Opt. 12, 064034 (2007).
[CrossRef]

Leuenberger, A.

A. Leuenberger, "Treatment of diabetic retinopathy with photocoagulation," Mod. Probl. Ophthalmol. 10, 564-576 (1972).
[PubMed]

Lin, C. P.

H. Lee, C. Alt, C. M. Pitsillides, and C. P. Lin, "Optical detection of intracellular cavitation during selective laser targeting of the retinal pigment epithelium: dependence of cell death mechanism on pulse duration," J. Biomed. Opt. 12, 064034 (2007).
[CrossRef]

R. Brinkmann, N. Koop, M. Oezdemir, C. Alt, G. Schuele, C. P. Lin, and R. Birngruber, "Targeting of the retinal pigment epithelium (RPE) by means of a rapidly scanned continuous wave (CW) laser beam," Lasers Surg. Med. 32, 252-264 (2003).
[CrossRef] [PubMed]

R. Brinkmann, G. Hüttmann, J. Rögener, J. Roider, R. Birngruber, and C. P. Lin, "Origin of RPE-cell damage by pulsed laser irradiance in the ns to µs time regime," Lasers Surg. Med. 27, 451-464 (2000).
[CrossRef] [PubMed]

C. P. Lin and M. W. Kelly, "Ultrafast time-resolved imaging of stress transient and cavitation from short pulsed laser irradiated melanin particles," Proc. SPIE 2391A, 294-299 (1995).
[CrossRef]

Loeffler, K. U.

M. Saham, K. U. Loeffler, P. Seifer, and M. Spitznas, "Effect of ultraviolet radiation on melanogenesis in four different types of cultured bovine ocular pigmented cells," Graefe's Arch. Clin. Exp. Ophthalmol. 239, 302-309 (2001).
[CrossRef]

Mieler, W. F.

B. L. Seagle, E. M. Gasyna, W. F. Mieler, and J. R. J. Norris, "Photoprotection of human retinal pigment epithelium cells against blue light-induced apoptosis by melanin free radicals from Sepia officinalis," Proc. Natl. Acad. Sci. U. S. A. 103, 16644-16648 (2006).
[CrossRef] [PubMed]

Norris, J. R. J.

B. L. Seagle, E. M. Gasyna, W. F. Mieler, and J. R. J. Norris, "Photoprotection of human retinal pigment epithelium cells against blue light-induced apoptosis by melanin free radicals from Sepia officinalis," Proc. Natl. Acad. Sci. U. S. A. 103, 16644-16648 (2006).
[CrossRef] [PubMed]

Oezdemir, M.

R. Brinkmann, N. Koop, M. Oezdemir, C. Alt, G. Schuele, C. P. Lin, and R. Birngruber, "Targeting of the retinal pigment epithelium (RPE) by means of a rapidly scanned continuous wave (CW) laser beam," Lasers Surg. Med. 32, 252-264 (2003).
[CrossRef] [PubMed]

Parrish, J. R.

R. R. Anderson and J. R. Parrish, "Selective photothermolysis: precise microsurgery by selective absorption of pulsed radiation," Science 220, 524-527 (1983).
[CrossRef] [PubMed]

Peisch, R. D.

S. Y. Schmidt and R. D. Peisch, "Melanin concentration in normal human retinal pigment epithelium," Invest. Ophthalmol. Vis. Sci. 27, 1063-1067 (1986).
[PubMed]

Pitsillides, C. M.

H. Lee, C. Alt, C. M. Pitsillides, and C. P. Lin, "Optical detection of intracellular cavitation during selective laser targeting of the retinal pigment epithelium: dependence of cell death mechanism on pulse duration," J. Biomed. Opt. 12, 064034 (2007).
[CrossRef]

Proulx, S.

S. Proulx, S. L. Guerin, and C. Salesse, "Effect of quiescence on integrin alpha5beta1 expression in human retinal pigment epithelium," Mol. Vis. 3, 473-481 (2003).

Putkey, F. R.

K. C. Dunn, A. E. Aotaki-Keen, F. R. Putkey, and L. M. Hjelmeland, "ARPE-19, A human retinal pigment epitheial cell line with differentiated properties," Exp. Eye Res. 62, 155-169 (1996).
[CrossRef] [PubMed]

Rögener, J.

R. Brinkmann, G. Hüttmann, J. Rögener, J. Roider, R. Birngruber, and C. P. Lin, "Origin of RPE-cell damage by pulsed laser irradiance in the ns to µs time regime," Lasers Surg. Med. 27, 451-464 (2000).
[CrossRef] [PubMed]

Roider, J.

R. Brinkmann, J. Roider, and R. Birngruber, "Selective retina therapy (SRT): a review on methods, techniques, preclinical and first clinical results," Bull. Soc. Belge. Ophtalmol. 302, 51-69 (2006).

R. Brinkmann, G. Hüttmann, J. Rögener, J. Roider, R. Birngruber, and C. P. Lin, "Origin of RPE-cell damage by pulsed laser irradiance in the ns to µs time regime," Lasers Surg. Med. 27, 451-464 (2000).
[CrossRef] [PubMed]

J. Roider, F. Hillenkamp, T. J. Flotte, and R. Birngruber, "Microphotocoagulation: Selective effects of repetitive short laser pulses," Proc. Natl. Acad. Sci. U.S.A. 90, 8643-8647 (1993).
[CrossRef] [PubMed]

Saham, M.

M. Saham, K. U. Loeffler, P. Seifer, and M. Spitznas, "Effect of ultraviolet radiation on melanogenesis in four different types of cultured bovine ocular pigmented cells," Graefe's Arch. Clin. Exp. Ophthalmol. 239, 302-309 (2001).
[CrossRef]

Salesse, C.

S. Proulx, S. L. Guerin, and C. Salesse, "Effect of quiescence on integrin alpha5beta1 expression in human retinal pigment epithelium," Mol. Vis. 3, 473-481 (2003).

Schmidt, S. Y.

S. Y. Schmidt and R. D. Peisch, "Melanin concentration in normal human retinal pigment epithelium," Invest. Ophthalmol. Vis. Sci. 27, 1063-1067 (1986).
[PubMed]

Schuele, G.

R. Brinkmann, N. Koop, M. Oezdemir, C. Alt, G. Schuele, C. P. Lin, and R. Birngruber, "Targeting of the retinal pigment epithelium (RPE) by means of a rapidly scanned continuous wave (CW) laser beam," Lasers Surg. Med. 32, 252-264 (2003).
[CrossRef] [PubMed]

Seagle, B. L.

B. L. Seagle, E. M. Gasyna, W. F. Mieler, and J. R. J. Norris, "Photoprotection of human retinal pigment epithelium cells against blue light-induced apoptosis by melanin free radicals from Sepia officinalis," Proc. Natl. Acad. Sci. U. S. A. 103, 16644-16648 (2006).
[CrossRef] [PubMed]

Seifer, P.

M. Saham, K. U. Loeffler, P. Seifer, and M. Spitznas, "Effect of ultraviolet radiation on melanogenesis in four different types of cultured bovine ocular pigmented cells," Graefe's Arch. Clin. Exp. Ophthalmol. 239, 302-309 (2001).
[CrossRef]

Spitznas, M.

M. Saham, K. U. Loeffler, P. Seifer, and M. Spitznas, "Effect of ultraviolet radiation on melanogenesis in four different types of cultured bovine ocular pigmented cells," Graefe's Arch. Clin. Exp. Ophthalmol. 239, 302-309 (2001).
[CrossRef]

Bull. Soc. Belge. Ophtalmol.

R. Brinkmann, J. Roider, and R. Birngruber, "Selective retina therapy (SRT): a review on methods, techniques, preclinical and first clinical results," Bull. Soc. Belge. Ophtalmol. 302, 51-69 (2006).

Exp. Eye Res.

K. C. Dunn, A. E. Aotaki-Keen, F. R. Putkey, and L. M. Hjelmeland, "ARPE-19, A human retinal pigment epitheial cell line with differentiated properties," Exp. Eye Res. 62, 155-169 (1996).
[CrossRef] [PubMed]

Graefe's Arch. Clin. Exp. Ophthalmol.

M. Saham, K. U. Loeffler, P. Seifer, and M. Spitznas, "Effect of ultraviolet radiation on melanogenesis in four different types of cultured bovine ocular pigmented cells," Graefe's Arch. Clin. Exp. Ophthalmol. 239, 302-309 (2001).
[CrossRef]

Health Phys.

R. Birngruber, F. Hillenkamp, and V. P. Gabel, "Theoretical investigations of laser thermal retinal injury," Health Phys. 48, 781-796 (1985).
[CrossRef] [PubMed]

Invest. Ophthalmol. Vis. Sci.

S. Y. Schmidt and R. D. Peisch, "Melanin concentration in normal human retinal pigment epithelium," Invest. Ophthalmol. Vis. Sci. 27, 1063-1067 (1986).
[PubMed]

J. Biomed. Opt.

H. Lee, C. Alt, C. M. Pitsillides, and C. P. Lin, "Optical detection of intracellular cavitation during selective laser targeting of the retinal pigment epithelium: dependence of cell death mechanism on pulse duration," J. Biomed. Opt. 12, 064034 (2007).
[CrossRef]

Lasers Surg. Med.

R. Brinkmann, G. Hüttmann, J. Rögener, J. Roider, R. Birngruber, and C. P. Lin, "Origin of RPE-cell damage by pulsed laser irradiance in the ns to µs time regime," Lasers Surg. Med. 27, 451-464 (2000).
[CrossRef] [PubMed]

R. Brinkmann, N. Koop, M. Oezdemir, C. Alt, G. Schuele, C. P. Lin, and R. Birngruber, "Targeting of the retinal pigment epithelium (RPE) by means of a rapidly scanned continuous wave (CW) laser beam," Lasers Surg. Med. 32, 252-264 (2003).
[CrossRef] [PubMed]

Mod. Probl. Ophthalmol.

A. Leuenberger, "Treatment of diabetic retinopathy with photocoagulation," Mod. Probl. Ophthalmol. 10, 564-576 (1972).
[PubMed]

Mol. Vis.

S. Proulx, S. L. Guerin, and C. Salesse, "Effect of quiescence on integrin alpha5beta1 expression in human retinal pigment epithelium," Mol. Vis. 3, 473-481 (2003).

Proc. Natl. Acad. Sci. U. S. A.

B. L. Seagle, E. M. Gasyna, W. F. Mieler, and J. R. J. Norris, "Photoprotection of human retinal pigment epithelium cells against blue light-induced apoptosis by melanin free radicals from Sepia officinalis," Proc. Natl. Acad. Sci. U. S. A. 103, 16644-16648 (2006).
[CrossRef] [PubMed]

Proc. Natl. Acad. Sci. U.S.A.

J. Roider, F. Hillenkamp, T. J. Flotte, and R. Birngruber, "Microphotocoagulation: Selective effects of repetitive short laser pulses," Proc. Natl. Acad. Sci. U.S.A. 90, 8643-8647 (1993).
[CrossRef] [PubMed]

Proc. SPIE

C. P. Lin and M. W. Kelly, "Ultrafast time-resolved imaging of stress transient and cavitation from short pulsed laser irradiated melanin particles," Proc. SPIE 2391A, 294-299 (1995).
[CrossRef]

Science

R. R. Anderson and J. R. Parrish, "Selective photothermolysis: precise microsurgery by selective absorption of pulsed radiation," Science 220, 524-527 (1983).
[CrossRef] [PubMed]

Other

V. P. Gabel, R. Birngruber, and F. Hillenkamp, Visible and near infrared light absorption in pigment epithelium and choroids. In International Congress Series No.450, XXIII concilium Ophthalmologicum, K. Shimizu, ed.Princeton, NJ: Excerpta Medica:1978:658-662). (Excerpta Medica, Princeton, NJ, 1978).
[PubMed]

M. F. Marmor and T. J. Wolfensberger, The retinal pigment epithelium (Oxford University Press, New York, 1998).

A. E. Siegman, Defining and Measuring Laser Beam Parameters. In Laser Beam Characterization (Sociedad Espanola de Optica, Madrid, 1993).

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Figures (6)

Fig. 1.
Fig. 1.

Schematic representation of the experimental setup. Laser irradiation was reflected by a mirror to the stage where RPE cells were seated. The slit lamp was used to capture the video images of laser irradiation spots. Laser irradiation was performed under visual control.

Fig. 2.
Fig. 2.

Pulsed dye laser profile analysis. (a) Light micrograph of laser spot. Slight asymmetry is caused by imaging optics; (b) Gaussian laser intensity profile analyzed by MATLAB 6.5 (arbitrary units). Scale bar 500 µm.

Fig. 3.
Fig. 3.

Phase contrast microscope images of confluent ARPE-19 cells following 20 hrs of incubation with melanin. (a) 0.0 mg melanin/mg protein. (b) 0.03 mg melanin/mg protein. (c) 0.05 mg melanin/mg protein. Scale bar is 100µm.

Fig. 4.
Fig. 4.

Effects of laser irradiation on viability of pigmented RPE cells. Cells were stained with the viable dye calcein (green) and dead stain ethidium homodimer-1 (EthD-1) (red). A) Laser radiant exposure at 5 mJ/cm2. B) Laser radiant exposure at 55 mJ/cm2. C) Laser radiant exposure at 700 mJ/cm2. Melanin content is 0.05mg/mg protein. Scale bar is 400 µm. The light and dark brown spots are the reflection of grid edges. D) Effects of laser irradiation on viability of non-pigmented RPE cells at laser radiant exposure of 2550 mJ/cm2. Scale bar is 640 µm.

Fig. 5.
Fig. 5.

Viability of pigmented RPE cells as a function of laser radiant exposure and melanin content. The lines represent the least-square linear fits of the experimental data with the following regression parameters (a=slope; b=ordinate; threshold=intersection with 100% viability line; r=regression coefficient). Melanin content 0.03 mg/mg protein: a=0.99; b=-9.8×10-4; r=0.98; threshold=1140 mJ/cm2 (Corresponding to 50% viability); Melanin content 0.05mg/mg protein: a=1.05; b=-1.1×10-3; r=0.98; threshold=460 mJ/cm2 (Corresponding to 50% viability). P values shown are comparisons with non-pigmented cells.***: p<0.001. The meanings of symbols are as following. Solid circle: experimental data for 0.0 mg/mg protein melanin content; solid square: experimental data for 0.03 mg/mg protein melanin content; solid triangle: experimental data for 0.05 mg/mg protein melanin content.

Fig. 6.
Fig. 6.

Selective targeting of pigmented retinal pigment epithelium cells at a laser radiant exposure of 1100 mJ/cm2. (a) Fluorescence image (red nuclear staining represents dead cells, green cytoplasmic staining represents live cells); (b) Phase contrast image. Scale bar is 10 µm; (c) Viability of pigmented and non-pigmented RPE cells. ***: p<0.001. N=6.

Equations (1)

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I ( r , z ) = E ( r , z ) 2 2 η = I 0 ( w 0 w ( z ) ) 2 exp ( 2 r 2 w 2 ( z ) )

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