Title

Fluorene-based fluorescent probes with high two-photon action cross-sections for biological multiphoton imaging applications

Authors

Authors

K. J. Schafer-Hales; K. D. Belfield; S. Yao; P. K. Frederiksen; J. M. Hales;P. E. Kolattukudy

Comments

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Abbreviated Journal Title

J. Biomed. Opt.

Keywords

two-photon fluorescence; reactive dyes; multiphoton bioimaging; PHOTOPHYSICAL PROPERTIES; DERIVATIVES; MICROSCOPY; EXCITATION; DYES; FLUOROPHORES; ABSORPTION; DESIGN; SENSOR; SERIES; Biochemical Research Methods; Optics; Radiology, Nuclear Medicine &; Medical Imaging

Abstract

Two-photon fluorescence microscopy is a powerful tool for the study of dynamic cellular processes and live-cell imaging. Many commercially available fluorescent probes have been used in multiphoton-based imaging studies despite exhibiting relatively low two-photon absorption cross-section values in the tunability range of ultrafast Ti:sapphire lasers commonly used in multiphoton microscopy imaging. Furthermore, available fluorophores may be plagued with low fluorescence quantum yield and/or photoinstability (i.e., photobleaching) on exposure to the high peak power and photon density provided by the ultrafast laser source. To address the demand for better performing dyes, we prepare fluorophores tailored for multiphoton imaging. These fluorophores are based on the fluorene ring system, known to exhibit high fluorescence quantum yield (> 0.7) and high photostability. Furthermore, an amine-reactive fluorescent probe for the covalent attachment onto amine-containing biomolecules is also prepared. Epi-fluorescence and two-photon fluorescence microscopy images of H9c2 rat cardiomyoblasts stained with an efficient two-photon absorbing fluorene fluorophore is demonstrated. Additionally, single-photon spectral characteristics of the amine-reactive fluorophore, as well as the two-photon absorption cross sections of its model adduct in solution, and spectral characterization of a bovine serum albumin (BSA) as a model bioconjugate are presented. (c) 2005 Society of Photo-Optical Instrumentation Engineers.

Journal Title

Journal of Biomedical Optics

Volume

10

Issue/Number

5

Publication Date

1-1-2005

Document Type

Article; Proceedings Paper

Language

English

First Page

8

WOS Identifier

WOS:000233711300004

ISSN

1083-3668

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