Measurement of the relative non-degenerate two-photon absorption cross-section for fluorescence microscopy

Measurement of the relative non-degenerate two-photon absorption cross-section for fluorescence microscopy

In non-degenerate two-photon microscopy (ND-TPM), the required energy for fluorescence excitation occurs via absorption of two photons of different energies derived from two synchronized pulsed laser beams. ND-TPM is a promising imaging technology offering flexibility in the choice of the photon ene...

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Bibliographic Details
Published in:Optics express Vol. 27; no. 6; pp. 8335 - 8347
Main Authors: Sadegh, Sanaz, Yang, Mu-Han, Ferri, Christopher G L, Thunemann, Martin, Saisan, Payam A, Devor, Anna, Fainman, Yeshaiahu
Format: Journal Article
Language:English
Published: United States Optical Society of America 18-03-2019
Online Access:Get full text
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Summary:In non-degenerate two-photon microscopy (ND-TPM), the required energy for fluorescence excitation occurs via absorption of two photons of different energies derived from two synchronized pulsed laser beams. ND-TPM is a promising imaging technology offering flexibility in the choice of the photon energy for each beam. However, a formalism to quantify the efficiency of two-photon absorption (TPA) under non-degenerate excitation, relative to the resonant degenerate excitation, is missing. Here, we derive this formalism and experimentally validate our prediction for a common fluorophore, fluorescein. An accurate quantification of non-degenerate TPA is important to optimize the choice of photon energies for each fluorophore.
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These senior authors equally contributed to this study
These authors contributed equally to this work
ISSN:1094-4087
1094-4087
DOI:10.1364/OE.27.008335