Data Tables
Fluorescence Lifetime Standards
The table of lifetime standards provides you with lifetime data on standard fluorophores that have single-exponential decays. These data can be used to test your lifetime instrumentation for systematic errors. For convenience we have divided them into nanosecond and picosecond standards.
Nanosecond Lifetime Standards | Lifetime [ns] | Conditions for Lifetime Measurement | Excitation [nm] | Emission [nm] | Ref. |
---|---|---|---|---|---|
NADH | 0.4 | 0.1 M PB 7.4, 20°C | 330-370 | 400-600 | 1 |
NATA | 3.0 | 0.1 M PB 7.0, 20°C | 275 | 310-400 | 1 |
p-Terphenyl | 1.05 | Ethanol | 280-320 | 310-412 | 2 |
PPD | 1.20 | Ethanol | 240-340 | 310-440 | 2 |
PPO | 1.4 | Ethanol | 280-350 | 330-480 | 2 |
POPOP | 1.35 | Ethanol Abs. | 280-390 | 370-540 | 2 |
Dimethyl-POPOP | 1.45 | Ethanol | 300-400 | 390-560 | 2 |
2-Aminopurine | 11.34 | Water | 290 | 380 | 2 |
L-Tyrosine | 3.27 | Water | 285 | 300 | 2 |
Anthranilic Acid | 8.6 | Water | 290 | 400 | 2 |
Indole | 4.49 | Water | 290 | 360 | 2 |
Fluorescein, dianion | 4.1±0.1 | NaOH/Water | 400 | 490-520 | 3 |
Rhodamine B | 1.74±0.02 | Water, 20°C | 400 | 583 | 4 |
PB = phosphate buffer
NATA = N-Acetyl-L-tryptophanamide
PPD = 1.5-diphenyl-1,3,4-oxadiazole
PPO = 2.5-diphenyl-oxazole
POPOP = 1, 4-bis(5-phenyloxazole-2-yl)benzene
Picosecond Lifetime Standards | Lifetime [ns] | Conditions for Lifetime Measurement | Excitation [nm] | Emission [nm] | Ref. |
---|---|---|---|---|---|
DMS | 0.880 | Cyclohexane, 25°C | 280-375 | 375-475 | 2 |
DFS | 0.328 | Cyclohexane, 25°C | 280-375 | 375-450 | 2 |
DBS | 0.176 | Cyclohexane, 25°C | 280-385 | 375-475 | 2 |
DCS | 0.066 | Cyclohexane, 25°C | 280-420 | 300-500 | 2 |
Rose Bengal | 0.519 | Methanol, 25°C | 556 | 572 | 2 |
DMS = 4-dimethylamino-4-methoxystilbene
DFS = 4-dimethylamino-4-fluorostilbene
DBS = 4-dimethylamino-4-bromostilbene
DCS = 4-dimethylamino-4-cyanostilbene
References
- J.R. Lakowicz, Principles of Fluorescence Spectroscopy, 1st Ed., Plenum Press, New York, London, 1983.
- J.R. Lakowicz, Principles of Fluorescence Spectroscopy, 2nd Ed., Kluwer Academic/Plenum Publishers, New York, London, Moscow, Dordrecht, 1999.
- D. Magde, G.E. Rojas, and P. Seybold, Solvent Dependence of the Fluorescence Lifetimes of Xanthene Dyes. Photochem. Photobiol. 70, 737, 1999.
- Boens, N., Qin, W., Basaric, N., Hofkens, J., Ameloot, M., Pouget, J., Lefevre, J-P., Valeur, B., Gratton, E., vandeVen, M., Silva, N.D., Jr., Engelborghs, Y., Willaert, K., Sillen, A., Rumbles, G., Phillips, D., Visser, A.J.W.G., van Hoek, A., Lakowicz, J.R., Malak, H., Gryczynski, I., Szabo, A.G., Krajcarski, D.T., Tamai, N., Miura, A., Analytical Chemistry, 79(5), p. 2137-2149
All of the following parameters and more may influence the measured lifetime value. Check values against literature, and see above references for more detail. Temperature, buffer, presence of oxygen (a fluorescence quencher), concentration, purity of the fluorophore dye, quality of equipment used (including cuvettes and optical filters)