I have a 50-50 mixture of a gemini surfactant and Tween 80 , I have not been able to determine the number of aggregation fluorescence , pyrene and CTB with no quencher and 9 -methyl anthracene and ruthenium either, how could I do?
Are you sure that micelles are formed in this system? Have you determined the cmc?The aggregation number is not an easy property to determine by static fluorescence, because the method is based on a considerable range of assumptions. If you have access to such an instrument it is better to use (static) light scattering.
The best fluorescence option for Nag determination is TRFQ (time-resolved fluorescence quenching) with pyrene. It is not trivial, but works well for many systems (see e.g. Almgren et al. J Colloid Int. Sci. 81, 486 (1981) as a simple introduction to the method).
If static fluorescence is your only option, then I am afraid that there is no guarantee that any fluorophore will work for your system. For example, I could not use pyrene to obtain the Nag of a zwitterionic surfactant and I got terrible results with coumarin 153 on the system, while both dyes work with SDS.
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