I am trying to calibrate the focal volume of FCS setup by using Alexa 488 dye. The experiments is being carried out at room temperature (25 degrees). I have to take the diffusion coefficient of Alexa to find out the volume. But in some papers it is 465 and in some literature values it is taken as 220. If I take the hydrodynamic radius of Alexa to be 5.7 A and calculate it then the value is 430.
Now I am confused as to which value to actually consider.
Any help is appreciated.
(I have 488nm laser, hence using Alexa 488)
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