I am using a Beckr and Hickl Tau 130 TCSPC setup for fluorescence lifetime measurement of different fluorophores. For excitation, I am using a pulsed laser (440 nm). For now, I am using fluorescein dianion as my standard but I would like to try other standards as well just to see if my TCSPC setup is giving right results. So, I am looking for a fluorophore which absorbs light at 440 nm and have a stable lifetime. Thank you!
Saurabh,
Here are some possibilities:
Coumarin 153 in methanol, excitation wavelength 295-442 nm, lifetime 4.3 ns.
reference: Boens, et al. (2007) Anal. Chem. 79: 2137-2149.
Coumarin 540 in acetonitrile, absorption maximum 454 nm, lifetime 3.2 ns.
reference: Jones, et al. (1985) J. Phys. Chem. 89: 294-300.
Coumarin 535 in ethanol, absorption maximum 433 nm, lifetime 3.2 ns
reference:
DCM in acetonitrile, absorption maximum 460 nm, lifetime 1.9 ns.
reference: Bourson and Valeur (1989) J. Phys. Chem. 93, 3871-3876.
Perylene in cyclohexane, absorption maximum 436 nm, lifetime 4.7 ns.
reference: Berlman (1965) Handbook of Fluorescence Spectra, Academic Press.
Auramine O in 1-decanol, absorption maximum 435 nm, lifetime 92 ps.
reference: Gautam and Harriman (1994) J. Chem. Soc., Faraday Trans. 90, 697-701.
Acridine orange in water, absorption maximum 450 nm, lifetime 1.7 ns.
reference: Kubota and Steiner (1977) Biophys. Chem. 6: 279-289.
I hope this some help.
Cheers,
John
Coumarin is a popular dye that absorbs at wavelengths around 400nm. See, http://dx.doi.org/10.1021/ac062160k .
Saurabh,
Here are some possibilities:
Coumarin 153 in methanol, excitation wavelength 295-442 nm, lifetime 4.3 ns.
reference: Boens, et al. (2007) Anal. Chem. 79: 2137-2149.
Coumarin 540 in acetonitrile, absorption maximum 454 nm, lifetime 3.2 ns.
reference: Jones, et al. (1985) J. Phys. Chem. 89: 294-300.
Coumarin 535 in ethanol, absorption maximum 433 nm, lifetime 3.2 ns
reference:
DCM in acetonitrile, absorption maximum 460 nm, lifetime 1.9 ns.
reference: Bourson and Valeur (1989) J. Phys. Chem. 93, 3871-3876.
Perylene in cyclohexane, absorption maximum 436 nm, lifetime 4.7 ns.
reference: Berlman (1965) Handbook of Fluorescence Spectra, Academic Press.
Auramine O in 1-decanol, absorption maximum 435 nm, lifetime 92 ps.
reference: Gautam and Harriman (1994) J. Chem. Soc., Faraday Trans. 90, 697-701.
Acridine orange in water, absorption maximum 450 nm, lifetime 1.7 ns.
reference: Kubota and Steiner (1977) Biophys. Chem. 6: 279-289.
I hope this some help.
Cheers,
John
Hey Christoph!
Thanks for your reply! The two fluorophores that you mentioned; do they absorb light at 440nm? Because that is my limitation. Thanks,
Saurabh
Hello Saurabh,
With all the above mentioned excellent suggestions I would like to add the name of flavin a adenine dinucleotide (FAD) in water.
absorption maximum at 375 nm and 440 nm, collection wavelength at 530 nm. You should be able to observe the following components at 25C
10 ps, 200 ps, 2.2 ns and 4.2 ns components
where the ultrafast 10 ps component dominates due to dominant stack conformation in water or buffer.
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