Hello everybody
I need a piece of advice on this topic:
I am working with oil in water droplets decorated with a fluorescent tag and ssDNA
To avoid droplets' coalescence I need to add a surfactant in the water, the SDS (0.5-1%) works well in that.... the back of the coin is that it acts as a quencher on my fluorescent tag.
does anybody of you have experience in this phenomenon?
thank you in advance
cheers
The quenching may be due to the negative charge on the SDS if your fluorescent tag has a positive charge. The problem may be reduced, in that case, by switching to a different detergent, one that is either positively charged or neutral.
Adam B Shapiro thank you
I am using an AlexaFluor streptavidin tag, that reacts with the biotinylated-lipid on the surface of the droplets.
I will try with a nonionic surfactant like Tween80 at a really low concentration to avoid that it creates micelle by itself!
(its cmc is equal to 0.012mM)
Alexa Fluor 488 is probably the most commonly used Alexa Fluor. It has both positive and negative charges. The longer wavelength Alexa Fluors also have both positive charge and negative charges.
Another possible reason for the quenching could be a hydrophobic interaction between the detergent and dye. If that is the reason, changing the detergent probably won't do much good because all detergents have hydrophobic parts, by definition.
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