BRG Photophysics & Photochemistry


Biophotonics & Bioimaging


Photophysics & Photochemistry


High-density Optical Data Storage


Functional Materials

One vs Two-photon Excitation

In 1931, it was predicted theoretically by Maria Goppert-Mayer[1] that photons of lesser energy together can cause an excitation "normally" produced by the absorption of a single photon of higher energy in a process called two-photon excitation. If the excited molecule is fluorescent, it can emit a single photon of fluorescence as if it were excited by a single higher energy photon (see photos below). Since its first demonstration by Webb et al in 1990, two-photon microscopy has been applied to a variety of imaging tasks and has now become the technique of choice for fluorescence microscopy in thick tissue and in live animals[2]. The Z-scan technique pioneered by Prof. Eric W. Van Stryland and Prof. David J. Hagan is the standard method to measure nonlinear absorption[3].

References:

[1] M. Goppert-Mayer, Ann. Phys., 1931, 9, 273-294.

[2] W. R. Zipfel, R. M. Williams, W. W. Webb, Nat. Biotechnol., 2003, 21, 1369-1377.

[3] M. Sheik-Bahea, A. A. Said, T. H. Wei, D. J. Hagan, E. W. Van Stryland, IEEE J. Quantum Electronics, 1990, 26, 760-769.

Below is a demonstration performed in our laboratory that show the high localization of two-photon excitation.


Experimental description

The light from a tunable Coherent Mira 900 Ti:Sapphire laser (tuned to 760 nm, 200 fs pulse width, 76 MHz repetition rate) was equally split into two beams. A second-harmonic generator (APE, Germany) was used to double the frequency of one beam, which was used as the one-photon excitation source. The fundamental and second-harmonic beams were both expanded to be about 12 mm in diameter and then focused into a 1cm quartz cuvette by two 10x (UPLANFLN 10x/0.30, Olympus) objective lenses in opposite directions. Dye with a concentration of 5*10-5 M was pre-loaded inside the cuvette.


Results

 

 

Publications

  1. Ghazvini Zadeh, E. H.; Bondar, M. V.; Mikhailov, I. A.; Belfield, K. D. “Linear Photophysics, Stimulated Emission and Ultrafast Spectroscopy of New Two-Photon Absorbing Diketopyrrolopyrrole Derivatives” Journal of Physical Chemistry C 2015, 119, 8864-8875.

  2. Irfan, M.; Belfield, K. D.; Saeed, A. “Carbazole/Fluorene Based Conjugated Small Molecules: Synthesis and Comparative Studies on the Optical, Thermal and Electrochemical Properties” RSC Advances 2015, 5, in press.

  3. Yue, X.; Armijo, Z.; King, K.; Bondar, M. V.; Morales, A. R.; Frazer, A.; Mikhailov, I. A.; Przhonska, O. V.; Belfield, K. D. “Steady-state and femtosecond transient absorption spectroscopy of new two-photon absorbing fluorene-containing quinolizinium cation membrane probes” ACS Applied Materials and Interfaces 2015, 7, 2833-2846.

  4. Belfield. K. D.; Bondar, M. V.; Yao, S.; Mikhailov, I. A.; Polikanov, V. S.; Przhonska, O. V.  “Femtosecond Spectroscopy of Superfluorescent Fluorenyl Benzothiadiazoles with Large Two-Photon and Excited State Absorption” Journal of Physical Chemistry C 2014, 118, 13790-13800. DOI: 10.1021/jp503106k

  5. Bashmakova, N.V.; Shaydyuk, Y.O.; Levchenko, S.M.; Masunov, A.E.; Przhonska, O.V.; Bricks, J.L.; Kachkovsky, O.D.; Slominsky, Y.L.; Piryatinski, Y.P.; Belfield, K.D.; Bondar, M.V. “Design and Electronic Structure of New Styryl Dye Bases: Steady-State and Time-Resolved Spectroscopic Studies.” Journal of Physical Chemistry A 2014, 118, 4502-4509.

  6. Moreshead, W. V.; Przhonska, O. V.; Bondar, M. V.; Kachkovski, A. D.; Nayyar, I. H.; Masonov, A. E.; Woodward, A. W.; Belfield, K. D. “Design of a New Optical Materials with Broad Spectrum Linear and Two-photon Absorption and Solvatochromism” Journal of Physical Chemistry C 2013, 117, 23133-23147.

  7. Belfield, K. D.; Bondar, M. V.; Haniff, H. S.; Mikhailov, I. A.; Luchita, G.; Przhonska, O. V. “Superfluorescent Squaraine with Efficient Two-photon Absorption and High Photostability” ChemPhysChem, 2013, 14, 3532-3542.

  8. Belfield, K. D.; Bondar, M. V.; Morales, A. R.; Frazer, A. F.; Mikhailov, I.; Przhonska, O. V. “Photophysical Properties and Ultrafast Excited State Dynamics of a New Two-Photon Absorbing Thiopyranyl Probe” Journal of Physical Chemistry C 2013, 117, 11941-11952.

  9. Yue, X.; Yanez, C. O.; Yao, S.; Belfield, K. D. “Selective Cell Death by Photochemically Induced pH Imbalance in Cancer Cells” Journal of the American Chemical Society 2013, 135, 2112-2115.

  10. Morales, A. R.; Frazer, A.; Woodward, A. W.; Ahn-White, H.-Y.; Fonari, A.; Tongwa, P.; Timofeeva, T. V.; Belfield, K. D. “Design, Synthesis, Structural and Spectroscopic Studies of Push-Pull Two-Photon Absorbing Chromophores with Acceptor Groups of Varying Strength” Journal of Organic Chemistry 2013, 78, 1014-1025.

  11. Zhao, K.; Liu, T.; Wang, G.; Chang, X.; Xue, D.; Belfield, K. D.; Fang, Y. “A Butterfly-Shaped Pyrene Derivative of Cholesterol and its uses as a Fluorescent Probe” Journal of Physical Chemistry B, 2013, 117, 5659-5667.

  12. Belfield, K. D.; Bondar, M. V.; Morales, A. R.; Yue, X.; Luchita, G.; Przhonska, O. V.; Kachkovsky, O. D. “Two-Photon Absorption and Time-Resolved Stimulated Emission Depletion Spectroscopy of a New Fluorenyl Derivative” ChemPhysChem 2012, 13, 3481-3491.

  13. Belfield, K. D.; Bondar, M. V.; Morales, A. R.; Yue, X.; Luchita, G.; Przhonska, O. V. “Transient Excited-state Absorption and Gain Spectroscopy of a Two-Photon Absorbing Probe with Efficient Superfluorescent Properties” Journal of Physical Chemistry C 2012, 116, 11261-11271.

  14. Belfield, K. D.; Bondar, M. V.; Morales, A. R.; Padhila, L. A.; Przhonska, O. V.; Wang, X. “Two-photon STED Spectral Determination for a New V-shaped Organic Fluorescent Probe with Efficient Two-photon Absorption” ChemPhysChem 2011, 12, 2755-2762.

  15. Luchita, G.; Bondar. M. V.; Yao, S.; Mikhaylov, I. A.; Yanez, C. O.; Przhonska, O. V.; Masunov, A. E.; Belfield, K. D. “Efficient Photochromic Transformation of a New Fluorenyl Diarylethene: One- and Two-photon Absorption Spectroscopy” ACS Applied Materials and Interfaces 2011, 3, 3559-3567.

  16. Belfield, K. D.; Bondar, M. V.; Hernandez, F. E.; Przhonska, O. V.; Wang, X.; Yao, S. “Superfluorescent Fluorenyl Probe with Efficient Two-photon Absorption” Physical Chemistry Chemical Physics 2011, 13, 4303-4310.

  17. Parthasarathy, A.; Ahn, H.-Y.; Belfield, K. D.; Schanze, K. S. “Two-photon Excited Florescence of a Conjugated Polyelectrolyte and its Application in Cell Imaging” ACS Applied Materials and Interfaces 2010, 2, 2744-2748. DOI: 10.1021/am100784m

  18. Belfield, K. D.; Andrade, C. D.; Yanez, C. O.; Bondar, M. V.; Hernandez, F. E.; Przhonska, O. V. “New Two-Photon Absorbing Probe with Efficient Superfluorescent Properties” Journal of Physical Chemistry B 2010, 114, 14087-14095. DOI: 10.1021/jp107343k

  19. Belfield, K. D.; Bondar, M. V.; Yanez, C. O.; Hernandez, F. E.; Przhonska, O. V. “One- and Two-photon Stimulated Emission Depletion of a Sulfonyl-containing Fluorene Derivative” Journal of Physical Chemistry B 2009, 113, 7101-7106. http://pubs.acs.org/doi/full/10.1021/jp902060m

  20. Morales, A. M.; Schafer-Hales, K. J.; Yanez, C. O.; Bondar, M. V.; Przhonska, O. V.; Markus, A. I.; Belfield, K. D. “Excited State Intramolecular Proton Transfer and Photophysics of a New Fluorenyl Two-Photon Fluorescent Probe” ChemPhysChem 2009, 10, 2073-2081. http://dx.doi.org/10.1002/cphc.200900032

  21. Belfield, K. D.; Bondar, M. V.; Yanez, C. O.; Hernandez, F. E.; Przhonska, O. V. Two-photon Absorption and Lasing Properties of New Fluorene Derivatives” Journal of Materials Chemistry 2009, 19, 7498-7502. http://dx.doi.org/10.1039/b820950b

  22. Belfield, K. D.; Bondar, M. V.; Hernandez, F. E.; Masunov, A. E.; Mikhailov, I. A.; Morales, A. R.; Przhonska, O. V.; Yao, S. “Two-photon Absorption Properties of New Fluorene-based Singlet Oxygen Photosensitizers” Journal of Physical Chemistry C 2009, 113, 4706-4711. http://dx.doi.org/10.1021/jp8102832

  23. Rodriguez, L.; Ahn, H.-Y.; Belfield, K. D. “Femtosecond Two-photon Absorption Measurements Based on the Accumulative Photo-thermal Effect and the Rayleigh Interferometer” Optics Express 2009, 17, 19617-19628.

  24. Bondar, M. V. Przhonska, O. V. Yanez, C. O. Belfield, K. D. “New Fluorene Molecules with Efficient Two-photon Absorption for Multidisciplinary Nonlinear Optical Applications” Ukranian Journal of Physics 2009, 54, 14-21.

  25. Belfield, K. D.; Bondar, M. V.; Hernandez, F. E.; Przhonska, O. V. “Photophysical Characterization, Two-Photon Absorption and Optical Power Limiting of Two Fluorenylperylene Diimides” Journal of Physical Chemistry C 2008, 112, 5618-5622. http://dx.doi.org/10.1021/jp711950z

  26. Balu, M.; Padhila, L. A.; Hagan, D. J.; Van Stryland, E. W.; Yao, S.; Belfield, K. D.; Zheng, S. J.; Barlow, S.; Marder, S. “Broadband Z-scan characterization using a high-spectral-irradiance, high-quality supercontinuum” Journal of the Optical Society of America B – Optical Physics 2008, 25, 159-165.

  27. Belfield, K. D.; Bondar, M. V.; Hernandez, F. E.; Przhonska, O. V.; Yao, S. “Two-Photon Absorption Cross Section Determination for Fluorene Derivatives: Analysis of the Methodology and Elucidation of the Origin of the Absorption Processes” Journal of Physical Chemistry B 2007, 111, 12723-12729. http://dx.doi.org/10.1021/jp074456f

  28. Belfield, K. D.; Bondar, M. V.; Kachkovsky, O. D.; Przhonska, O. V.; Yao, S. “Solvent effect on the steady-state fluorescence anisotropy of two-photon absorbing fluorene derivatives” Journal of Luminescence 2007, 126 (1), 14-20. http://www.sciencedirect.com/science/article/pii/S0022231306004637?np=y

  29. Andrasik, S. J.; Belfield, K. D.; Bondar, M. V.; Hernandez, F. E.; Morales, A. R.; Przhonska, O. V.; Yao, S. “One- and Two-Photon Singlet Oxygen Generation with New Fluorene-based Photosensitizers“ ChemPhysChem 2007, 8, 399-404. http://dx.doi.org/10.1002/cphc.200600568