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

Raz Gvishi; Garry Berkovic; Zvi Kotler; P. Krief; J. Y. Becker; Vladimir Khodorkovsky

doi:10.1117/12.424574

24 April 2001 New long-wave and highly efficient two-photon fluorophores for multiphoton microscopy

Raz Gvishi, Garry Berkovic, Zvi Kotler, P. Krief, J. Y. Becker, Vladimir Khodorkovsky

Proceedings Volume 4262, Multiphoton Microscopy in the Biomedical Sciences; (2001) https://doi.org/10.1117/12.424574
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

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-n-A-n-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: (1) Very large two-photon absorption coefficients (6 > 1000 GM); (2) Peak TP excitation wavelength which are strongly shifted to the red ((lambda) 1 micrometer); (3) Large fluorescence quantum efficiency; (4) 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.

Citation Download Citation

Raz Gvishi, Garry Berkovic, Zvi Kotler, P. Krief, J. Y. Becker, and Vladimir Khodorkovsky "New long-wave and highly efficient two-photon fluorophores for multiphoton microscopy", Proc. SPIE 4262, Multiphoton Microscopy in the Biomedical Sciences, (24 April 2001); https://doi.org/10.1117/12.424574

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