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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

  1. J.R. Lakowicz, Principles of Fluorescence Spectroscopy, 1st Ed., Plenum Press, New York, London, 1983.
  2. J.R. Lakowicz, Principles of Fluorescence Spectroscopy, 2nd Ed., Kluwer Academic/Plenum Publishers, New York, London, Moscow, Dordrecht, 1999.
  3. D. Magde, G.E. Rojas, and P. Seybold, Solvent Dependence of the Fluorescence Lifetimes of Xanthene Dyes. Photochem. Photobiol. 70, 737, 1999.
  4. 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)

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