Special Section on Optical Diagnostic Imaging from Bench to Bedside Spectroscopy

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

[+] Author Affiliations
Katherine J. Schafer-Hales

University of Central Florida, Department of Chemistry and Burnett College of Biomedical Sciences, 4000 Central Florida Boulevard, Orlando, Florida 32816

Kevin D. Belfield

University of Central Florida, Department of Chemistry and College of Optics and Photonics: Center for Research and Education in Optics and Lasers and Florida Photonics Center of Excellence and Burnett College of Biomedical Sciences, 4000 Central Florida Boulevard, Orlando, Florida 32816

Sheng Yao

University of Central Florida, Department of Chemistry, 4000 Central Florida Boulevard, Orlando, Florida 32816

Peter K. Frederiksen

University of Central Florida, Department of Chemistry and College of Optics and Photonics: Center for Research and Education in Optics and Lasers and Florida Photonics Center of Excellence, 4000 Central Florida Boulevard, Orlando, Florida 32816

Joel M. Hales

University of Central Florida, College of Optics and Photonics: Center for Research and Education in Optics and Lasers and Florida Photonics Center of Excellence, 4000 Central Florida Boulevard, Orlando, Florida 32816

Pappachan E. Kolattukudy

University of Central Florida, Burnett College of Biomedical Sciences, 4000 Central Florida Boulevard, Orlando, Florida 32816

J. Biomed. Opt. 10(5), 051402 (October 17, 2005). doi:10.1117/1.2104528
History: Received February 04, 2005; Accepted March 10, 2005; Published October 17, 2005
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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.

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© 2005 Society of Photo-Optical Instrumentation Engineers

Citation

Katherine J. Schafer-Hales ; Kevin D. Belfield ; Sheng Yao ; Peter K. Frederiksen ; Joel M. Hales, et al.
"Fluorene-based fluorescent probes with high two-photon action cross-sections for biological multiphoton imaging applications", J. Biomed. Opt. 10(5), 051402 (October 17, 2005). ; http://dx.doi.org/10.1117/1.2104528


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