Abstract
We observed the steady-state and time-resolved emission of tetraphenylbutadiene (TPB) whea excited by simultaneous absorption of two photons (514 to 610 nm). The intensity initially increased quadratically with laser power, as expected for a two-photon process. At higher laser powers the intensity increases in TPB were subquadratic. The intensity and anisotropy decay times of TPB were unchanged under the locally intense illumination. Importantly, the time zero anisotropy of TPB was decreased under conditions where the intensity was subquadratic. Furthermore, the subquadratic dependence on incident power was not observed for two-photon excitation of 2,5-diphenyloxazole (PPO), for which the incident wavelength does not overlap with the emission spectrum. These results are consistent with stimulated emission (light quenching) of TPB at high laser intensities. The phenomenon of light quenching may be important for other fluorophores used in biochemical research, particularly for the high local intensities used for two-photon excitation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
J. R. Lakowicz, I. Gryczynski, Z. Gryczynski, E. Danielsen, and M. J. Wirth (1992)J. Phys. Chem. 96, 3000–3006.
J. R. Lakowicz, I. Gryczynski, E. Danielsen, and J. K. Frisoli (1992)Chem. Phys. Lett. 194, 282–287.
A. A. Rehms and P. R. Callis (1993)Chem. Phys. Lett., in press.
J. R. Lakowicz and I. Gryczynski (1992)Biophys. Chem. 45, 1–6.
J. R. Lakowicz, I. Gryczynski, and E. Danielsen (1992)Chem. Phys. Lett. 191, 47–53.
J. R. Lakowicz and I. Gryczynski (1992)J. Fluoresc. 2, 117–121.
H. Szmacinski, I. Gryczynski, and J. R. Lakowicz (1993)Photochem. Photobiol. 58, 341–345.
W. M. McClain (1971)J. Chem. Phys. 55, 2789–2796.
A. P. Aleksandrov, V. I. Bredikhin, and V. N. Genkin (1971)Sov. Phys. JETP 33, 1078–1082.
W. M. McClain (1971)J. Chem. Phys. 55, 2789–2796.
M. J. Wirth, A. Koskelo, and M. J. Sanders (1981)Appl. Spectrosc. 35, 14–21.
P. R. Callis (1993) J. Chem. Phys., in press.
W. Denk, J. H. Strickler, and W. W. Webb (1990)Science 248, 73–76.
W. W. Webb (1990) inMICRO 90, IOP, London, pp. 445–450.
J. R. Lakowicz, H. Szmacinski, K. Nowaczyk, and M. L. Johnson (1992)Proc. Natl. Acad. Sci. 89, 1271–1275.
J. R. Lakowicz, H. Szmacinski, K. Nowaczyk, K. W. Bendi and M. L. Johnson (1992)Anal. Biochem. 202, 316–330.
D. W. Piston, D. R. Sandison, and W. W. Webb (1992)Proc. SPIE 1640, 379–388.
A. Ansari and A. Szabo (1993)Biophys. J. 64, 838–851.
M. D. Galanin, B. P. Kirsanov, and Z. A. Chizhikova (1969)Sov. Phys. JETP Lett. 9, 502–507.
J. R. Lakowicz, V. Bogdanov, J. Kuśba, and I. Gryczynski (1994) in preparation.
J. R. Lakowicz, I. Gryczynski, V. Bogdanov, and J. Kuśba (1994)J. Phys. Chem. 98, 334–342.
I. Gryczynski, V. Bogdanov, and J. R. Lakowicz (1993) sub mitted for publication.
O. Svelto (1989) in D. C. Hanna (Trans. and Ed.),Principles of Lasers, 3rd ed., Plenum Press, New York, p. 52.
K. I. Rudik, L. G. Pikulik, L. P. Senkevich, M. Y. Kostko, and A. I. Maksimov (1976)Zh. Prikladnoi Spektrosk.25, 450–454 (translated).
Y. T. Mazurenko (1973)Opt. Spectrosc. 35, 137–139.
A. I. Butko, E. S. Voropai, V. A. Gaisenok, V. A. Saechnikov, and A. M. Sarzhevskii (1982)Opt. Spectrosc. 52, 153–156.
J. R. Lakowicz and B. P. Maliwal (1985)Biophys. Chem. 21, 61–78.
J. R. Lakowicz, G. Laczko, and I. Gryczynski (1986)Rev. Sci. Instrum. 57, 2499–2506.
G. Laczko, J. R. Lakowicz, I. Gryczynski, Z. Gryczynski, and H. Malak (1990)Rev. Sci. Instrum. 61, 2331–2337.
J. R. Lakowicz, H. Cherek, and A. Balter (1981)J. Biochem. Biophys. Methods 5, 131–146.
J. R. Lakowicz and I. Gryczynski (1993)Biophys. Chem.,47, 1–7.
A. I. Butko, E. S. Voropai, I. I. Zholnerevich, V. A. Saechnikov, and A. M. Sarzhevskii (1978)Invest. Akad. Nauk SSSR Ser. Fiz. 42, 626–630 (English translation, pp. 150–153).
P. Peretti and P. Ranson (1971)Opt. Comm. 3, 62–64.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gryczynski, I., Bogdanov, V. & Lakowicz, J.R. Light quenching of tetraphenylbutadiene fluorescence observed during two-photon excitation. J Fluoresc 3, 85–92 (1993). https://doi.org/10.1007/BF00865322
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00865322