Simultaneous Two-Photon Activation of Type-I Photodynamic Therapy Agents


  • †AMT, 4′-aminomethyl-4,5,8-trimethylpsoralen; CW, continuous wave; DMSO, dimethyl sulfoxide; EDTA, ethylenedi-aminetetraacetic acid; HpD, hematoporphyrin derivative; LIB, laser-induced breakdown; MNNG, /V-methyl-W-nitro-JV-nitrosoguan-idine; 8-MOP, 8-methoxypsoralen; 2-NF, 2-nitrofluorene; MVL, minimum visible lesion; NIR, near infrared; PDT, photodynamic therapy; Pheo-a, pheophorbide-a; TBE, Tris-borate-EDTA buffer; Ti:S, titanium: sapphire; TPE, two-photon excitation; Tris, tris(hydroxymethyl)aminomethane.

*Oak Ridge National Laboratory, Oak Ridge, TN 37831-6113, USA. Fax: 423-576-2078; e-mail:


The excitation and emission properties of several psoralen derivatives are compared using conventional single-photon excitation and simultaneous two-photon excitation (TPE). Two-photon excitation is effected using the output of a mode-locked titanium: sapphire laser, the near infrared output of which is used to promote non-resonant TPE directly. Specifically, the excitation spectra and excited-state properties of 8-methoxypsoralen and 4′-aminomethyl-4,5,8-trimethylpsoralen are shown to be equivalent using both modes of excitation. Further, in vitro feasibility of two-photon photodynamic therapy (PDT) is demonstrated using Salmonella typhimurium. Two-photon excitation may be beneficial in the practice of PDT because it would allow replacement of visible or UV excitation light with highly penetrating, nondamag-ing near infrared light and could provide a means for improving localization of therapy. Comparison of possible laser excitation sources for PDT reveals the titanium: sapphire laser to be exceptionally well suited for nonlinear excitation of PDT agents in biological systems due to its extremely short pulse width and high repetition rate that together provide efficient PDT activation and greatly reduced potential for biological damage