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Proceedings Paper

Evidence for excitation of fluorescence in RPE melanin by multiphoton absorption
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Paper Abstract

Previously, we reported that ultrashort, near infrared (NIR) laser pulses caused more DNA breakage in cultured retinal pigment epithelial (RPE) cells than did CW, NIR laser radiation delivering a similar radiant exposure. We hypothesized that this difference was due to multiphoton absorption in an intracellular chromophore such as the RPE melanin. We investigated two-photon excitation of fluorescence in a suspension of isolated bovine RPE melanosomes exposed to a 1-KHz train of approximately 50- fsec laser pulses at 810 nm from a Ti:Sapphire laser, and compared this to the fluorescence excited by CW exposures at 406 nm from a Krypton ion laser. Fluorescence was measured with a PC-based spectrometer. The CW sources excited fluorescence with a peak at 525 nm. The fluorescence intensity depended on the irradiance of the sample, as well as the melanosome concentration. Peak fluorescence was obtained with a suspension of ~2 x 107 melanin granules/ml. The 810-nm, ultrashort pulses also excited fluorescence, but with a broader, lower-amplitude peak. The weaker fluorescence signal excited by the 810-nm ultrashort pulse laser for a given melanosome concentration, compared to 406-nm CW excitation, is possibly due to the smaller two- photon absorption cross-section. These results indicate the involvement of multiphoton absorption in DNA damage.

Paper Details

Date Published: 27 June 2002
PDF: 8 pages
Proc. SPIE 4617, Laser Tissue Interaction XIII: Photochemical, Photothermal, and Photomechanical, (27 June 2002); doi: 10.1117/12.472521
Show Author Affiliations
Randolph D. Glickman, Univ. of Texas Health Science Ctr./San Antonio (United States)
Benjamin A. Rockwell, Air Force Research Lab. (United States)
Gary D. Noojin, Northrup Grumman Corp. (United States)
David J. Stolarski, Northrup Grumman Corp. (United States)
Michael L. Denton, Northrup Grumman Corp. (United States)

Published in SPIE Proceedings Vol. 4617:
Laser Tissue Interaction XIII: Photochemical, Photothermal, and Photomechanical
Steven L. Jacques; Donald Dean Duncan; Donald Dean Duncan; Sean J. Kirkpatrick; Sean J. Kirkpatrick; Andres Kriete; Andres Kriete, Editor(s)

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