I'm studying the interaction of different elements with fluorophore and have got interesting results so far, but it's difficult to understand what is happening in the reaction system. So is there any way out to know the possible mechanism?
Measuring fluorescence lifetimes will distinguish between static and dynamic quenching. Here is a good presentation on the subject:
https://www.chem.uzh.ch/dam/jcr:571775eb-8b0e-4930-8aa4-ad0da5c11aec/FluorescenceQuenching_HS16.pdf
Yes, I agree with Sir, Adam........Lifetime measurement is the most definitive way to distinguish between static and dynamic quenching. In case of dynamic quenching the quencher interacts with the excited state fluorophore and thus also affect the fluorescence lifetime of the sample. Whereas in case of static quenching there is complex formation between the fluorophore and quencher in the ground state and hence the fluorophores in their excited states do not get affected at all.
Another method is the temperature dependence measurement. The temperature effect on the quenching efficiency could be carefully examined to distinguish between these two mechanisms. Dynamic quenching efficiency is expected to increase with increase temperature due to larger diffusion coefficients at higher temperature, while increased temperature is likely to result in decreased stability of the complexes, and thus reduce the static quenching efficiency.
According to the valuable answer of Adam, please give us what a kind of fluorochrome (s) are you using and what a kind of quencher in your reaction system for quenching.
What a kind of complex can result in your rearch? Define it please.
In order to obtain valuables results and because lifetime measurements are very sensible to impurities, please note that your fluorochrome must be PURE and you must know the properties of it (p.e. Adam´s joined paper/ acridine orange/ Base? double salt or hydrochloride ? It is the same for the fluoresceine salt . P.e. the dilution of A.O. in solvent (concentration/ monomer/dimer ratio) plays a rolle too.
In order to understand the concentration quenching of fluorescence polarization ,
please obtain and read the following paper:
Th. Foerster, Ann. Phys. 2, 55, (1948) and related papers in references and
A. jablonski Bull. Acad. Polon. Sci, ser. sci. math.astr. et phys. 20,243, (1972).
Good luck
JRG
It purely depends on kind of system you are investigating, rest answer are given in previous replies.
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