Abstract
Two-photon-induced fluorescence is observed from the photodynamic phycobiliprotein phycoerythrin. Temporal, spectral, and intensity-dependent properties of the two-photon-induced fluorescence emission from phycoerythrin excited by a 1.06-mum laser beam are reported. The measured two-photon absorption cross section of phycoerythrin is an order of magnitude larger than that of Rhodamine 6G. The potential applications of phycobiliproteins for two-photon-induced fluorescence for microscopy of three-dimensional biological samples and three-dimensional optical memory are discussed.
Highlights
Two-photon-induced fluorescence has been a useful spectroscopic tool to elucidate the molecular electronic structure of excited states
Two-photon-induced fluorescence is a nonlinear process in which the fluorescence emission depends quadratically on the excitation laser intensity
As a result the fluorescence signal from twophoton excitation is dominated by the most intense region of the excitation beam. This nonlinear process provides an optical sectioning effect that allows for fluorescence imaging with 3-D resolution without using the confocal configuration
Summary
Two-photon-induced fluorescence has been a useful spectroscopic tool to elucidate the molecular electronic structure of excited states It has received considerable attention because of the potential for using two-photon-induced fluorescence for biomolecular imaging with three-dimensional3-Dresolution.[1,2] Two-photon-induced fluorescence is a nonlinear process in which the fluorescence emission depends quadratically on the excitation laser intensity. As a result the fluorescence signal from twophoton excitation is dominated by the most intense region of the excitation beam This nonlinear process provides an optical sectioning effect that allows for fluorescence imaging with 3-D resolution without using the confocal configuration. L. Kaplan were with the Biotechnology Division, U.S Army Natick Research, Development, and Engineering Center, Natick, Massachusetts 01760. D. Kaplan is with the Biotechnology Center, Tufts University, Medford, Massachusetts 02155.
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