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Simulation of the relationship between molecular weight (MW), rotational time, and FA. A: Graphic interpretation. B: Simulations are shown for dyes with various fluorescence lifetimes: 1 ns (cyanine dyes), 4 ns (fluorescein and Alexa Fluor 488), 20 ns (dansyl dyes, pyrens) and a hypothetical dye at 200 ns. Simulations assume ro fundamental anisotropy = 0.4 and rigid attachment of dyes to spherical carriers. C: Common classes of fluorophores and specific examples used in FA assays.
