New long wave and highly efficient two-photon fluorophores for multiphoton microscopy

R. Gvishi, G. Berkovic, Z. Kotler, P. Krief, J. Y. Becker, V. Khodorkovsky

Research output: Contribution to journalArticlepeer-review

6 Scopus citations


An important ingredient in improving Multi Photon Laser Scanning Microscopy, MPLSM, is the development of efficient exogenous two-photon fluorescent (TPF) probes. Here we report on a new class of two-photon fluorophores, specifically designed in order to maximize their efficiency in potential MPLSM applications. The fluorophores possess a symmetric Donor-Acceptor-Donor (D-π-A-π-D) structure with varying conjugation length and have strong donors and acceptors. We have studied the two-photon excitation (TPE) properties of these fluorophores and found the following properties: a) Very large two-photon absorption coefficients (δ > 1000GM); b) Peak TP excitation wavelength which are strongly shifted to the red (λ.∼ 1μm); c) Large fluorescence quantum efficiency; d) Large Stokes shifts of the fluorescence bands. These properties make them particularly suitable for imaging thicker samples, relying on the large improvement in TPE cross-sections and the reduced attenuation at both the excitation and emission wavelengths. We also describe TPE fluorescence anisotropy experiments revealing the tensorial shape of the fluorophores.

Original languageEnglish
Pages (from-to)340-346
Number of pages7
JournalProceedings of SPIE - The International Society for Optical Engineering
StatePublished - 1 Jan 2001


  • Conjugated molecules
  • D-A-D
  • Two-photon fluorophores


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