Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 to 1050 nm
Abstract
Measurements of two-photon fluorescence excitation (TPE) spectra are presented for 11 common molecular fluorophores in the excitation wavelength range 690 nm < λ < 1050 nm. Results of excitation by ∼100-fs pulses of a mode-locked Ti:sapphire laser are corroborated by single-mode cw Ti:sapphire excitation data in the range 710 nm < λ < 840 nm. Absolute values of the TPE cross section for Rhodamine B and Fluorescein are obtained by comparison with one-photon-excited fluorescence, assuming equal emission quantum efficiencies. TPE action cross sections for the other nine fluorophores are also determined. No differences between one-photon- and two-photon-excited fluorescence emission spectra are found. TPE emission spectra are independent of excitation wavelength. With both pulsed and cw excitation the fluorescence emission intensities are strictly proportional to the square of the excitation intensity to within ±4% for excitation intensities sufficiently below excited-state saturation.
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The current amplitudes showed a supralinear dependence on radiation power, with a power exponent of 2.5 ± 0.6 (mean ± s.d., n = 7).
“The current amplitudes showed a supralinear (power exponent, 2.5 ± 0.6, mean ± s.d., n = 7) dependence on the radiation power, as predicted (Fig. 1i) both by the two-photon excitation30 of MNI-glutamate and by the cooperativity (1.2–1.7) of glutamate binding to AMPA receptors31 (2pEPSCs were CNQX (6-cyano-7-nitroquinoxaline)-sensitive).”
From Dendritic spine geometry is critical for AMPA receptor expression in hippocampal CA1 pyramidal neurons (2001) · cites this paper as [30]
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- Authors
- Xu, C., & Webb, W. W.
- Affiliations
- Cornell University (US)
- ORCID
- Xu, Chris
- Venue
- Journal of the Optical Society of America B
- Publisher
- Optica Publishing Group
- Published
- 1996-03-01
- Volume
- 13
- Issue
- 3
- Pages
- 481
- ISSN
- 0740-3224 (ISSN-L)
- 1520-8540
- OpenAlex
- W2113504999
- Cited by
- 2,198
- References
- 0
- Citations by year
- 2026: 44 · 2025: 44 · 2024: 57 · 2023: 73 · 2022: 67 · 2021: 72 · 2020: 81 · 2019: 98 · 2018: 97 · 2017: 98 · 2016: 106 · 2015: 118 · 2014: 88 · 2013: 118 · 2012: 95
- FWCI
- 35.97
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of the Optical Society of America B (publishedVersion)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › Nonlinear Optical Materials Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Laser-Matter Interactions and Applications
- Keywords
- two-photon excitation, molecular fluorophores, rhodamine B, fluorescein, continuous-wave excitation
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